Cotton variety p07x.8212.rf

ABSTRACT

A cotton variety, designated P07X.8212.RF, the plants and seeds of the cotton variety P07X.8212.RF, methods for producing a cotton plant, either varietal or hybrid, produced by crossing the cotton variety P07X.8212.RF with itself or with another cotton plant, and hybrid cotton seeds and plants produced by crossing the variety P07X.8212.RF with another cotton variety or plant and to methods for producing a cotton plant containing in its genetic material one or more transgenes and to the transgenic cotton plants produced by that method. This disclosure also relates to cotton varieties derived from cotton variety P07X.8212.RF, to methods for producing other cotton varieties derived from cotton variety P07X.8212.RF, and to the varieties derived by the use of those methods.

This application claims the benefit of U.S. Provisional Application No.61/407,681 which was filed in the U.S. Patent and Trademark Office onOct. 28, 2010, the entire disclosure of which is hereby incorporated byreference in its entirety.

FIELD OF THE INVENTION

This invention is in the field of cotton breeding.

BACKGROUND OF THE INVENTION

Cotton (Gossypium spp.) is the world's most important textile fiber cropand is one of the world's most important oilseed crops. Cotton plantsprovide a source of human food, livestock feed, and raw material inindustry. Cotton seed is pressed for cooking oil and the residualcottonseed oil meal is used for animal feed. Industrial uses of cottoninclude candle wicks, twine, paper and a multitude of fabric products.

The genus Gossypium is very large, currently containing more than 50species. Two tetraploid species of Gossypium have spinnable seed fiberscalled lint. These two species are G. hirsutum (referred to as AmericanUpland cotton) and G. barbadense (referred to as Pima cotton).

The goal of a cotton breeder is to improve a cotton plant's performanceand therefore, its economic value by combining various desirable traitsinto a single plant. Improved performance is manifested in many ways.Higher yields of cotton plants contribute to increased lint fiberproduction, more profitable agriculture and lower cost of products forthe consumer. Improved plant health increases the yield and quality ofthe plant and reduces the need for application of protective chemicals.Adapting cotton plants to a wider range of production areas achievesimproved yield and vegetative growth. Improved plant uniformity enhancesthe farmer's ability to mechanically harvest cotton.

Cotton is a dicot plant with perfect flowers, i.e., cotton has male,pollen-producing organs and separate female, pollen receiving organs onthe same flower. The cultivated cotton flower is surrounded by threetriangular bracts forming what is commonly known as squares. The flowercontains an open corolla with five petals, a staminal column bearingclusters of stamens and forming a tube that encloses the style. Thecompound pistil consists of three to five carpels with stigmasprotruding above the anthers. The ovary develops into a three- tofive-loculed capsule or boll. From seven to nine seeds are set withineach lock or locule. On the day preceding anthesis, a twisted corollaemerges from the square. On the day of anthesis, the corolla opens andpollen shedding occurs. The corolla turns red the day following anthesisand later falls from the plant. Pollination occurs with the opening ofthe anthers and shedding of pollen on the stigma or with the deposit ofpollen on the stigma by insects.

Because cotton has both male and female organs on the same flower,cotton breeding techniques take advantage of the plant's ability to bebred by both self-pollination and cross-pollination. Self-pollinationoccurs when pollen from the male organ is transferred to a female organon the same flower on the same plant. Self-incompatibility is a form ofinfertility caused by the failure of cotton plants with normal pollenand ovules to set seed due to some physiological hindrance that preventsfertilization. Self-incompatibility restricts self-pollination andinbreeding and fosters cross-pollination. Cross-pollination occurs whenpollen from the male organ on the flower of one plant is transferred toa female organ on the flower on a different plant.

A plant is sib-pollinated (a type of cross-pollination) when individualswithin the same family or line are used for pollination (i.e. pollenfrom a family member plant is transferred to the stigmas of anotherfamily member plant). Self-pollination and sib-pollination techniquesare traditional forms of inbreeding used to develop new cottonvarieties, but other techniques exist to accomplish inbreeding. Newcotton varieties are developed by inbreeding heterozygous plants andpracticing selection for superior plants for several generations untilsubstantially homozygous plants are obtained. During the inbreedingprocess with cotton, the vigor of the lines decreases and after asufficient amount of inbreeding, additional inbreeding merely serves toincrease seed of the developed variety. Cotton varieties are typicallydeveloped for use in the production of hybrid cotton lines.

Natural, or open pollination, occurs in cotton when bees or otherinsects transfer pollen from the anthers to the stigmas and may includeboth self- and cross-pollination. Such pollination is accomplishedalmost entirely by the bees or other pollinating insects as the pollenis heavy and sticky and accordingly, interplant transfer of pollen bythe wind is of little importance. Vigor is restored when two differentvarieties are cross-pollinated to produce the first generation (F₁)progeny. A cross between two defined substantially homozygous cottonplant varieties always produces a uniform population of heterozygoushybrid cotton plants and such hybrid cotton plants are capable of beinggenerated indefinitely from the corresponding variety cotton seedsupply.

When two different, unrelated cotton parent plant varieties are crossedto produce an F₁ hybrid, one parent variety is designated as the male,or pollen parent, and the other parent variety is designated as thefemale, or seed parent. Because cotton plants are capable ofself-pollination, hybrid seed production requires elimination of orinactivation of pollen produced by the female parent to render thefemale parent plant male sterile. This serves to prevent the cottonplant variety designated as the female from self-pollinating. Differentoptions exist for controlling male fertility in cotton plants such asphysical emasculation, genetic male sterility, cytoplasmic malesterility and application of gametocides. Incomplete removal of maleparent plants from a hybrid seed production field before harvestprovides the potential for unwanted production of self-pollinated orsib-pollinated seed, which may be unintentionally harvested and packagedwith hybrid seed.

The development of new cotton plant varieties and hybrid cotton plantsis a slow, costly interrelated process that requires the expertise ofbreeders and many other specialists. The development of new varietiesand hybrid cotton plants in a cotton plant breeding program involvesnumerous steps, including: (1) selection of parent cotton plants(germplasm) for initial breeding crosses; (2) inbreeding of the selectedplants from the breeding crosses for several generations to produce aseries of varieties, which individually breed true and are highlyuniform; and (3) crossing a selected variety with an unrelated varietyto produce the F₁ hybrid progeny having restored vigor.

Cotton plant varieties and other sources of cotton germplasm are thefoundation material for all cotton breeding programs. Despite theexistence and availability of numerous cotton varieties and other sourcegermplasm, a continuing need still exists for the development ofimproved germplasm because existing parent cotton varieties lose theircommercial competitiveness over time. Embodiments of the presentdisclosure address this need by providing a novel cotton inbred varietydesignated P07X.8212.RF that possesses broad adaptation and excellentyield stability in the full-maturity cotton growing regions of the US;excellent fiber properties such as micronaire, length, strength (g/tex),and fiber uniformity; and the Roundup Ready® Flex transgenic event fortolerance to glyphosate herbicide. P07X.8212.RF also possesses toleranceto Fusarium oxysporum race 4, a serious pathogen on Pima cottons.P07X.8212.RF contributes such characteristics to hybrids relative toother similar hybrids in the same maturity groups. To protect and toenhance yield production, trait technologies and seed treatment optionsprovide additional crop plan flexibility and cost effective controlagainst insects, weeds and diseases, thereby further enhancing thepotential of this variety and hybrids with P07X.8212.RF as a parent.

SUMMARY OF THE INVENTION

Embodiments of this disclosure relate to a cotton variety designatedP07X.8212.RF that includes plants and seeds of cotton varietyP07X.8212.RF. Further embodiments relate to lint having novelcharacteristics whether or not produced by the claimed cotton variety.Methods for producing cotton plants, such as cotton plant varieties,hybrid cotton plants, or other cotton plants, as by crossing cottonvariety P07X.8212.RF with itself or any different cotton plant are anintegral part of certain embodiments, as are the resultant cotton plantsincluding the plant parts and seeds. Other embodiments further relate tomethods for producing P07X.8212.RF-derived cotton plants, to methods forproducing male sterile P07X.8212.RF cotton plants, e.g., cytoplasmicmale sterile P07X.8212.RF cotton plants and to methods for regeneratingsuch plants from tissue cultures of regenerable cells as well as theplants obtained therefrom. Methods for producing a cotton plantcontaining in its genetic material one or more transgenes, and thetransgenic cotton plants produced by that method, are also a part offurther embodiments.

In one embodiment, the present disclosure relates to a seed of thecotton variety designated P07X.8212.RF, or a part thereof,representative seed of the variety having been deposited under ATCCAccession No. PTA-11344. In a further aspect, the disclosure relates toa part of this seed, selected from the group consisting of hull(seedcoat), germ and endosperm. In a further aspect, the disclosurerelates to this seed, further comprising a coating. In a further aspect,the disclosure relates to a substantially homogenous composition of thisseed.

In another embodiment, the present disclosure relates to a method forproducing a seed of a cotton plant, comprising: (a) planting seed of thecotton variety designated P07X.8212.RF in proximity to itself or todifferent seed from a same variety; (b) growing plants from the seedunder pollinating conditions; and (c) harvesting the resultant seed. Ina further aspect, the disclosure relates to a cotton seed produced bythis method. In a further aspect, the disclosure relates to this method,further comprising pre-treating the seed before performing step (a). Ina further aspect, the disclosure relates to this method, furthercomprising treating the growing plants or soil surrounding the growingplants with an agricultural chemical.

In another embodiment, the present disclosure relates to a cotton plantproduced by growing a seed of the cotton variety designatedP07X.8212.RF. In a further aspect, the disclosure relates to a part ofthis cotton plant, selected from the group consisting of an intact plantcell, a plant protoplast, embryos, pollen, flowers, seeds, linters,fibers, pods, gossypol glands, leaves, bolls, stems, roots, root tips,and anthers. In a further aspect, the disclosure relates to fibers ofthis plant. In a further aspect, the disclosure relates to staples ofthis plant. In a further aspect, the disclosure relates to a cottonplant, or a part thereof, having all the physiological and morphologicalcharacteristics of this cotton plant. In a further aspect, thedisclosure relates to a substantially homogenous population of thesecotton plants. In a further aspect, the disclosure relates to thissubstantially homogenous population of cotton plants, wherein thepopulation is present in a field and the field further comprises other,different cotton plants.

In another embodiment, the present disclosure relates to a method forproducing a cotton plant, comprising: (a) crossing cotton variety plantP07X.8212.RF, representative seed of the cultivar having been depositedunder ATCC Accession No. PTA-11344, with another different cotton plantto yield progeny cotton seed. In a further aspectthe disclosure relatesto this method, wherein the other, different cotton plant is a cottonvariety. In a further aspect, the disclosure relates to this method,further comprising: (b) growing the progeny cotton seed from step (a)under self-pollinating or sib-pollinating conditions for about 5 toabout 7 generations; and (c) harvesting resultant seed. In a furtheraspect, the disclosure relates to this method, further comprisingselecting plants obtained from growing at least one generation of theprogeny cotton seed for a desirable trait.

In another embodiment, the present disclosure relates to a method ofintroducing a desired trait into cotton variety P07X.8212.RF,representative seed of the variety having been deposited under ATCCAccession No. PTA-11344, comprising: (a) crossing P07X.8212.RF plantswith plants of another cotton variety that comprise a desired trait toproduce F₁ progeny plants; (b) selecting F₁ progeny plants that have thedesired trait; (c) crossing selected progeny plants with P07X.8212.RFplants to produce backcross progeny plants; (d) selecting for backcrossprogeny plants that comprise the desired trait and physiological andmorphological characteristics of cotton variety P07X.8212.RF; and (e)performing steps (c) and (d) one or more times in succession to producethe selected or higher backcross progeny plants that comprise thedesired trait and all of the physiological and morphologicalcharacteristics of cotton variety P07X.8212.RF listed in Table 1 asdetermined at the 5% significance level when grown in the sameenvironmental conditions. In a further aspect, the disclosure relates tothis method, wherein the plants of the other cotton variety comprise adesired trait selected from the group consisting of male sterility,herbicide resistance, insect resistance, and resistance to bacterial,fungal and viral disease. In a further aspect, the disclosure relates tothis method, further comprising using direct or indirect selection todetermine whether the desired trait is present in a progeny plant.

In another embodiment, the present disclosure relates to a method forproducing a cotton plant, comprising: (a) crossing a cotton plantproduced by growing a seed of the cotton variety designated P07X.8212.RFwith another different cotton plant to produce a diploid or progenyplant; (b) generating a haploid progeny plant from the diploid progenyplant; (c) generating a diploid plant from the haploid progeny plant;and (d) selecting the diploid cotton plant. In a further aspect, thedisclosure relates to this method, wherein the haploid progeny plant isgenerated by culturing a haploid explant from the diploid progeny plant.In a further aspect, the disclosure relates to this method, wherein thehaploid progeny plant is generated by crossing the progeny plant withanother, different plant that induces haploid cotton plants. In afurther aspect, the disclosure relates to this method, wherein theother, different plant is a cotton plant that comprises ahaploid-inducing gene. In a further aspect, the disclosure relates tothis method, wherein the diploid plant of step (c) is generated bysubjecting the haploid progeny plant to a treatment that induceschromosome doubling in the cultured explant. In a further aspect, thedisclosure relates to this method, wherein the diploid plant of step (c)is generated by self-pollinating the haploid progeny plant.

In another embodiment, the present disclosure relates to a method forproducing a cotton plant, comprising: (a) inducing a mutation in acotton plant produced by growing a seed of the cotton variety designatedP07X.8212.RF, or a part thereof; and, (b) selecting mutated cottonplants. In a further aspect, the disclosure relates to this method,wherein the mutation is artificially induced by a method selected fromthe group consisting of elevated temperature, long-term seed storage,tissue culture conditions, radiation, and chemical mutagenesis.

In another embodiment, the present disclosure relates to a method forproducing a cotton plant variety, comprising: (a) growing firstgeneration hybrid cotton plants having P07X.8212.RF, representative seedof the variety having been deposited under ATCC Accession No. PTA-11344,as a parent cotton plant; (b) inbreeding the first generation hybridcotton plants or crossing the first generation hybrid cotton plants withdifferent cotton plants to yield progeny cotton seed; (c) growing theprogeny cotton seed of step (b) to yield further progeny cotton seed;(d) repeating the inbreeding or the crossing and the growing steps of(b) and (c) from about 0 to about 7 times to generate cotton varietalplants. In a further aspect, the disclosure relates to a cotton plantvariety produced by this method.

In another embodiment, the present disclosure relates to a method forproducing cotton variety P07X.8212.RF, representative seed of thevariety having been deposited under ATCC Accession No. PTA-11344,comprising: (a) planting a collection of seed comprising seed of ahybrid, one of whose parents is P07X.8212.RF, the collection alsocomprising seed of the variety P07X.8212.RF; (b) growing plants from thecollection of seed; (c) identifying a varietal parent plant; (d)controlling pollination in a manner that preserves the homozygosity ofthe varietal parent plant; and, (e) harvesting the resultant seed fromthe identified varietal parent plant which was pollinated to preserveits homozygosity. In a further aspect, the disclosure relates to thismethod, wherein step (c) comprises identifying plants with decreasedvigor. In a further aspect, the disclosure relates to a method forproducing a varietal cotton plant comprising: sib-pollinating plantsobtained by growing the harvested resultant seed of step (e) of thismethod. In a further aspect, the disclosure relates to a method forproducing a varietal cotton plant comprising: crossing P07X.8212.RFcotton plants with cotton plants obtained by growing the hybrid seed ofstep (a) of this method.

In another embodiment, the present disclosure relates to a method forproducing a hybrid cotton seed comprising crossing a first varietalparent cotton plant with a second varietal parent cotton plant andharvesting resultant hybrid cotton seed, wherein the first varietalcotton plant or the second varietal cotton plant is a cotton plantproduced by growing a seed of the cotton variety designatedP07X.8212.RF.

In another embodiment, the present disclosure relates to a method forproducing a hybrid cotton seed comprising the steps of: (a) planting inpollinating proximity seeds of a first and a second varietal parentcotton plants, wherein the first varietal cotton plant or the secondvarietal cotton plant is a cotton plant produced by growing a seed ofthe cotton variety designated P07X.8212.RF; (b) cultivating the seeds ofthe first and the second varietal cotton plants into plants that bearflowers; (c) controlling the male fertility of the first or the secondvarietal cotton plant to produce a male sterile cotton plant; (d)allowing cross-pollination to occur between the first and secondvarietal cotton plants; and, (e) harvesting seeds produced on the malesterile cotton plant. In a further aspect, the disclosure relates tothis method, wherein the varietal cotton plant that is the cotton plantproduced by growing a seed of the cotton variety designated P07X.8212.RFis a female parent. In a further aspect, the disclosure relates to thismethod, wherein the varietal cotton plant that is the cotton plantproduced by growing a seed of the cotton variety designated P07X.8212.RFis a male parent. In a further aspect, the disclosure relates to ahybrid cotton seed produced by this method. In a further aspect, thedisclosure relates to a hybrid cotton plant, or parts thereof, producingby growing this hybrid cotton seed. In a further aspect, the disclosurerelates to a tissue culture of regenerable cells from this hybrid cottonplant. In a further aspect, the disclosure relates to a cotton seedobtained by growing the hybrid cotton seed produced by this method andharvesting the resultant cotton seed from produced plants.

In another embodiment, the present disclosure relates to a method forproducing a hybrid cotton seed comprising crossing a first varietalparent cotton plant with a second varietal parent cotton plant andharvesting the resultant hybrid cotton seed, wherein the first varietalcotton plant or the second varietal cotton plant is a progeny plant of across of the cotton plant produced by growing a seed of the cottonvariety designated P07X.8212.RF and another varietal cotton plant. In afurther aspect, the disclosure relates to a hybrid cotton seed producedby this method. In a further aspect, the disclosure relates to a hybridcotton plant, or a part thereof, produced by growing this hybrid cottonseed. In a further aspect, the disclosure relates to a cotton seedproduced by growing this hybrid cotton plant and harvesting theresultant cotton seed.

In another embodiment, the present disclosure relates to an F₁ hybridseed produced by crossing the varietal cotton plant produced by growinga seed of the cotton variety designated P07X.8212.RF with another,different cotton plant. In a further aspect, the disclosure relates to ahybrid cotton plant, or a part thereof, produced by growing this hybridcotton seed. In a further aspect, the disclosure relates to this hybridcotton seed, wherein the other, different plant is not a member of thebarbadense species. In a further aspect, the disclosure relates to thishybrid cotton seed, wherein the other, different plant is a member ofthe hirsutum species. In a further aspect, the disclosure relates tothis hybrid cotton seed, wherein the other, different plant is a memberof a genus Gossypium. In a further aspect, the disclosure relates tothis hybrid cotton seed, wherein the other, different plant is a memberof the family Malvaceae.

In another embodiment, the present disclosure relates to a method forproducing a P07X.8212.RF-derived cotton plant, comprising: (a) crossingcotton variety P07X.8212.RF, representative seed of the variety havingbeen deposited under ATCC Accession No. PTA-11344, with a second cottonplant to yield progeny cotton seed; and (b) growing said progeny cottonseed, under plant growth conditions, to yield the P07X.8212.RF-derivedcotton plant. In a further aspect, the disclosure relates to aP07X.8212.RF-derived cotton plant, or a part thereof, produced by thismethod. In a further aspect, the disclosure relates to this method,further comprising: (c) crossing the P07X.8212.RF-derived cotton plantwith itself or another cotton plant to yield additionalP07X.8212.RF-derived progeny cotton seed; (d) growing the progeny cottonseed of step (c) under plant growth conditions, to yield additionalP07X.8212.RF-derived cotton plants; and (e) repeating the crossing andgrowing steps of (c) and (d) from 0 to 7 times to generate furtherP07X.8212.RF-derived cotton plants. In a further aspect, the disclosurerelates to this method, still further comprising utilizing plant tissueculture methods and/or haploid breeding to derive progeny of theP07X.8212.RF-derived cotton plant.

In another embodiment, the present disclosure relates to a tissueculture of regenerable cells from the cotton plant produced by growing aseed of the cotton variety designated P07X.8212.RF. In a further aspect,the disclosure relates to this tissue culture, the cells or protoplastsof the tissue culture being from a tissue selected from the groupconsisting of embryos, pollen, flowers, seeds, linters, fibers, pods,gossypol glands, leaves, bolls, stems, roots, root tips, and anthers. Ina further aspect, the disclosure relates to a cotton plant regeneratedfrom this tissue culture, wherein the regenerated plant expresses allthe morphological and physiological characteristics of varietyP07X.8212.RF.

In another embodiment, the present disclosure relates to a cotton plantwith all of the physiological and morphological characteristics ofcotton variety P07X.8212.RF, wherein the cotton plant is produced by atissue culture process using the cotton plant produced by growing a seedof the cotton variety designated P07X.8212.RF as a starting material forthe process.

In another embodiment, the present disclosure relates to a method forregenerating a cotton plant comprising the steps of: (a) culturing anexplant comprising a tissue selected from the group consisting of atissue obtained from cotton plant variety P07X.8212.RF, representativeseed having been deposited under ATCC Accession No. PTA-11344, animmature tissue obtained from a hybrid cotton plant having P07X.8212.RFas a parent, and a P07X.8212.RF-derived cotton plant; and, (b)initiating regeneration. In a further aspect, the disclosure relates tothis method, wherein the explant is an immature tissue.

In another embodiment, the present disclosure relates to a cotton plantproduced by growing a seed of the cotton variety designatedP07X.8212.RF, wherein the P07X.8212.RF plant is rendered male sterile.In a further aspect, the disclosure relates to this cotton plant,wherein the male sterile P07X.8212.RF plant is a cytoplasmic malesterile plant.

In another embodiment, the present disclosure relates to a method forproducing a male sterile P07X.8212.RF cotton plant, comprising: (a)crossing a varietal cotton plant produced by growing a seed of thecotton variety designated P07X.8212.RF, with a cytoplasmic male sterilecotton plant that generates haploids; (b) identifying haploid plants;and, (c) crossing the haploid plants with the varietal cotton plantP07X.8212.RF to produce male sterile P07X.8212.RF cotton plants.

In another embodiment, the present disclosure relates to a cotton plant,or a part thereof, produced by growing a seed of the cotton varietydesignated P07X.8212.RF, wherein the plant or part thereof has beentransformed so that its genetic material contains one or more transgenesoperably linked to one or more regulatory elements. In a further aspect,the disclosure relates to a method for producing a cotton plant thatcontains in its genetic material one or more transgenes, comprisingcrossing this cotton plant with either a second plant of another cottonvariety, or a non-transformed cotton plant of the variety P07X.8212.RF,so that the genetic material of the progeny that result from the crosscontains the transgene(s) operably linked to a regulatory element. In afurther aspect, the disclosure relates to a cotton plant, or a partthereof, produced by this method.

In another embodiment, the present disclosure relates to a cotton plantproduced by growing a seed of the cotton variety designatedP07X.8212.RF, or a part thereof, further comprising one or moretransgenes. In a further aspect, the disclosure relates to a seed ofthis plant. In a further aspect, the disclosure relates to this cottonplant, wherein the one or more transgenes comprise a gene conferringupon said cotton plant insect resistance, disease resistance or virusresistance. In a further aspect, the disclosure relates to this cottonplant, wherein the gene conferring upon the cotton plant insectresistance is a Bacillus thuringiensis gene.

In another embodiment, the present disclosure relates to a cotton plantproduced by growing a seed of the cotton variety designatedP07X.8212.RF, or a part thereof, wherein the plant or a parts thereofhas been transformed so that its genetic material contains one or moretransgenes operably linked to one or more regulatory elements. In afurther aspect, the disclosure relates to this cotton plant, wherein theone or more transgenes comprise a gene conferring upon the cotton planttolerance to a herbicide. In a further aspect, the disclosure relates tothis cotton plant, wherein the herbicide is glyphosate, glufosinate, asulfonylurea or an imidazolinone herbicide, a hydroxyphenylpyruvatedioxygenase inhibitor or a protoporphyrinogen oxidase inhibitor.

In another embodiment, the present disclosure relates to a method forproducing a population of P07X.8212.RF progeny cotton plants comprising:(a) obtaining a first generation progeny cotton seed comprising theplant produced by growing a seed of the cotton variety designatedP07X.8212.RF as a parent; (b) growing the first generation progenycotton seed to produce F₁ generation cotton plants and obtaining self orsib pollinated seed from the F₁ generation cotton plants; and (c)producing successive filial generations to obtain a population ofP07X.8212.RF progeny cotton plants. In a further aspect, the disclosurerelates to the population of P07X.8212.RF progeny cotton plants producedby this method, the population, on average, deriving 50% of its allelesfrom P07X.8212.RF.

In another embodiment, the present disclosure relates to lint havingsubstantially the same characteristics of the lint produced by cottonvariety designated P07X.8212.RF, representative seed of the varietyhaving been deposited under ATCC Accession No. PTA-11344.

DETAILED DESCRIPTION OF THE INVENTION

I. Definitions of Plant Characteristics

In the description and examples that follow, a number of terms are used.To provide a clear and consistent understanding of the specification andclaims, including the scope to be given such terms, the followingdefinitions are provided.

Area(s) of Adaptation: This represents whether the cotton plant isadapted (A), not adapted (NA) or not tested (NT) for the followingareas: Eastern, Delta, Central, Blacklands, Plains, Western, Arizona,and San Joaquin Valley.

Plant Habit: This represents the general growth habit of the plant ratedas spreading, intermediate or compact.

Foliage: This represents the general appearance of the plant leavesrated as sparse, intermediate, or dense.

Stem Lodging: This represents the general appearance of the plant stemsrelative to their normal near vertical orientation rated as lodging,intermediate, or erect.

Fruiting Branch: This represents fruiting pattern rated as clustered,short, or normal.

Growth: This represents the growing pattern of the cotton plantfollowing a fruiting cycle rated as determinate, i.e., a completeinterruption of growth following a fruiting cycle, or indeterminate,i.e., a growth pattern in which stems continue to grow indefinitely.

Leaf Color: This represents a visual assessment of the leaf color of thecotton plant rated as greenish yellow, light green, medium green, darkgreen.

Boll Shape: This represents the shape of the boll rated as length lessthan width, length equal to width, or length more than width.

Boll Breadth: This represents a comparison of the boll width at itsmiddle and its base rated as broadest at base, or broadest at middle.

cm to 1st Fruiting Branch: This represents the distance between thecotyledonary node to the first fruiting branch in centimeters.

No. of Nodes to 1st Fruiting Branch: This represents the number of nodesfrom the cotyledonary node to the first fruiting branch, excluding thecotyledonary node.

Mature Plant Height: This represents the height in centimeters of thecotton plant from the cotyledonary node to terminal

Leaf Type: This represents the shape of the uppermost fully expandedleaf rated as normal, sub okra, okra, or super okra.

Leaf Pubescence: This represents the density of leaf trichomes (“hairs”)on the bottom surface excluding veins of the uppermost fully expandedleaf rated as absent, sparse, medium, or dense in terms oftrichomes/cm².

Stem Pubescence: This represents whether the stem pubescence isglabrous, intermediate, or hairy.

Leaf Glands: This represents the density of gossypol glands rated asabsent, sparse, normal, or more than normal.

Stem Glands: This represents the density of gossypol glands rated asabsent, sparse, normal, or more than normal.

Calyx Lobe: This represents the gossypol gland density on the calyx loberated as absent (normal), sparse, or more than normal.

Petal Color: This represents a visual assessment of the petal colorrated as cream or yellow.

Pollen Color: This represents a visual assessment of pollen color ratedas cream or yellow.

Petal Spot: This represents whether petal spot is present or absent onthe flowers of the cotton plant.

Seed Index: This represents the weight of 100 seeds in grams on a fuzzybasis.

Lint: This represents the fibers produced by a cotton plant that areassociated with the seed coat and may include either staples or linters.

Lint Index: This represents the weight of lint per 100 seeds in grams.

Number of Seeds per Boll: This represents the average number of seedsper boll on the cotton plant.

Grams Seed Cotton per Boll: This represents the average number of gramsof seed cotton per boll on the cotton plant.

Fiber Length: This represents fiber length expressed in hundredths of aninch as measured by High Volume Instrumentation (HVI).

Fiber Uniformity: This represents the uniformity of fiber length in asample as measured on the HVI, expressed as a percentage.

Fiber Strength: This represents the force required to rupture or tobreak a bundle of fibers as measured in grams per tex on the HVI.

Fiber Elongation: This represents the amount that a fiber sample willstretch before breakage and is a measure of the deformation of thecotton fiber at rupture expressed as percent change in length based onthe original fiber length as measured by HVI.

Fiber Micronaire: This represents a measure of the fineness of thefiber. Within a cotton cultivar, micronaire is also a measure ofmaturity. Micronaire differences are governed by changes in perimeter orin cell wall thickness, or by changes in both. Within a variety, cottonperimeter is fairly constant and maturity will cause a change inmicronaire. Consequently, micronaire has a high correlation withmaturity within a variety of cotton. Maturity is the degree ofdevelopment of cell wall thickness. Micronaire may not have a goodcorrelation with maturity between varieties of cotton having differentfiber perimeter. Micronaire values range from about 2.0 to 6.0 and havethe following meanings: below 2.9 very fine possible small perimeter butmature (good fiber), or large perimeter but immature (bad fiber); from2.9 to 3.7 fine various degrees of maturity and/or perimeter; 3.8 to 4.6average degree of maturity and/or perimeter; 4.7 to 5.5 coarse usuallyfully developed (mature), but larger perimeter; and 5.6 or greater verycoarse fully developed, large-perimeter fiber.

II. Cotton Variety P07X.8212.RF

-   -   A. Cotton Plant P07X.8212.RF

In accordance with one aspect of the present disclosure, provided is anew Pima (Gossypium barbadense) cotton seed and plants thereofdesignated P07X.8212.RF. Further embodiments relate to a method forproducing cotton seeds that includes, but is not limited to, the stepsof planting seed of cotton variety P07X.8212.RF in proximity to itselfor to different seed from a same family or line, growing the resultingcotton plants under self-pollinating or sib-pollinating conditions withadequate isolation, and harvesting resultant seed obtained from suchplants using techniques standard in the agricultural arts such as wouldbe necessary to bulk-up seed such as for hybrid production. Embodimentsof the present disclosure also relate to varietal seed produced by sucha method.

In any cross between cotton plant variety P07X.8212.RF and anothercotton plant variety, P07X.8212.RF may be designated as the male (pollenparent) or the female (seed parent). Optionally, the seed of cottonvariety P07X.8212.RF may be pre-treated to increase resistance of theseed and/or seedlings to stressed conditions, and further, the cottonplants or surrounding soil may be treated with one or more agriculturalchemicals before harvest. Such agricultural chemicals may includeherbicides, insecticides, pesticides and the like. Embodiments of thepresent disclosure also relate to a cotton plant that expressessubstantially all of the physiological and morphological characteristicsof cotton plant variety P07X.8212.RF and to a substantially homogenouspopulation of cotton plants having all the physiological andmorphological characteristics of cotton plant variety P07X.8212.RF. Anycotton plants produced from cotton plant variety P07X.8212.RF arecontemplated by embodiments of the present disclosure and are,therefore, within the scope thereof. A description of physiological andmorphological characteristics of cotton plant P07X.8212.RF is presentedin Table 1.

TABLE 1 Physiological and Morphological Characteristics of VarietyP07X.8212.RF Characteristic Value^(a) Area(s) of Adaptation Pima growingregions of CA, AZ, NM, and TX. Plant Habit Spreading Foliage Normal StemLodging Semi-erect Fruiting Branch Normal Growth Indeterminate LeafColor Green Boll Shape Normal Distance to 1st Fruiting Branch (cm) 27.7Nodes to 1st Fruiting Branch (number) 6.3 Mature Plant Height (cm) 89.2Leaf Type Normal Leaf Pubescence Semi-smooth Leaf Nectaries Present StemPubescence Semi-smooth Leaf Glands Present Stem Glands Present PetalColor Yellow Pollen Color Yellow Petal Spot (present or absent) PresentSeed Index (weight of 100 seeds in grams) 13.7 Lint Index (weight of 100seeds in grams) 8.7 Lint Percent 39.9 Gin Turnout 32.5 Fiber Length(hundredths of an inch) 145 Fiber Uniformity (percentage) 89.0 FiberStrength (grams per tex) 46.9 Fiber Elongation, E1 (percentage) 6.14Fiber Micronaire 4.09 Fiber Fineness 2.98 Fusarium Wilt Tolerant (race4) Verticillium Wilt Moderately Tolerant Root-Knot Nematode N/A^(a)Typical values, which may vary due to the environment, are listed.Other values that are substantially equivalent are within the scope ofthis invention.

It should be appreciated by one having ordinary skill in the art that,for the quantitative characteristics identified in Table 1, the valuespresented are typical values. These values may vary due to theenvironment and accordingly, other values that are substantiallyequivalent are also within the scope of embodiments of the disclosure.

Cotton variety P07X.8212.RF shows uniformity and stability within thelimits of environmental influence for the traits described in Table 1.Variety P07X.8212.RF has been self-pollinated a sufficient number ofgenerations with careful attention paid to uniformity of plant type toensure the homozygosity and phenotypic stability necessary to use inlarge scale, commercial production. The line has been increased both byhand and sib-pollinated in isolated fields with continued observationsfor uniformity. No variant traits have been observed or are expected inP07X.8212.RF.

Embodiments of the present disclosure also relate to one or more cottonplant parts of cotton plant P07X.8212.RF. Cotton plant parts includeplant cells, plant protoplasts, plant cell tissue cultures from whichcotton plants can be regenerated, plant DNA, plant calli, plant clumps,and plant cells that are intact in plants or parts of plants, such asembryos, ovules, pollen, stigmas, flowers, petals, seeds, bolls,gossypol glands, stems, leaves, fibers, roots, root tips, and the like.

-   -   B. Cotton Seed Designated P07X.8212.RF

A cotton seed is composed of three structural parts: (1) the pericarp,which is a protective outer covering (also known as bran or hull); (2)the germ (also known as an embryo); and (3) the endosperm. Anotheraspect of the present disclosure is one or more parts of cotton seedP07X.8212.RF, such as the pericarp of cotton seed P07X.8212.RF or thegerm and/or the endosperm of cotton seed P07X.8212.RF, which remain uponremoval of the pericarp and adhering remnants of the seed coat.

Cotton seed designated P07X.8212.RF may be provided as a substantiallyhomogenous composition of cotton seed designated P07X.8212.RF, that is,a composition that consists essentially of cotton seed P07X.8212.RF.Such a substantially homogenous composition of cotton seed P07X.8212.RFis substantially free from significant numbers of other varietal and/orhybrid seed so that the varietal seed forms from about 90% to about 100%of the total seed. Preferably, a substantially homogenous composition ofthe varietal cotton seed contains from about 98.5%, 99%, or 99.5% toabout 100% of the varietal seed, as measured by seed grow outs. Thesubstantially homogenous composition of varietal cotton seed ofembodiments of the disclosure may be separately grown to providesubstantially homogenous populations of varietal cotton plants. However,even if a population of varietal cotton plants is present in a fieldwith other different cotton plants, such as in a commercialseed-production field of single-cross hybrid cotton planted in a ratioof 1 male pollinator row to 4 female seed-parent rows, such a populationwould still be considered to be within the scope of embodiments of thepresent disclosure.

Cotton yield is affected by the conditions to which seeds and seedlings(young plants grown from seeds) are exposed. Seeds and seedlings may beexposed to one of, or a combination of, for example, cold, drought,salt, heat, pollutants, and disease, all of which are conditions thatpotentially retard or prevent the growth of crops therefrom. Forexample, temperature extremes are typical in the United States.Furthermore, diseases evolved from pathogens and deterioration caused byfungi are potentially harmful to seeds and seedlings. Thus, it isdesirable to treat seeds as by coating or impregnating the seeds withcompositions that render the seeds and seedlings grown therefrom morehardy when exposed to such adverse conditions.

Accordingly, another aspect of the present disclosure relates to acoated and/or impregnated seed or cotton variety designated P07X.8212.RFand to coated and/or impregnated seed derived therefrom. Various agentshave been used to treat seeds to increase resistance of the plants tostressed conditions, such as cold, drought, salt, and fungi. Such agentsinclude, for example, sodium methylphenyl-pentadienate, trichloroaceticacid, polyoxyalkylene-organo-siloxane block copolymer, 5-aminolevulinicacid, salicylic acid, thiamethoxam, potassium chloride, and polyvinylalcohol and are useful alone, or in combination in embodiments of thepresent disclosure.

When pre-treating seeds according to the present disclosure such asbefore the seeds are planted, the seeds are contacted with thecomposition of interest, as by coating seeds, spraying seeds, andsoaking seeds or a combination thereof, by methods well known to thoseskilled in the art.

-   -   C. Deposit Information

Applicants have made a deposit of at least 2,500 seeds of cotton varietyP07X.8212.RF with the American Type Culture Collection (ATCC), Manassas,Va. 20110 USA, under ATCC Accession No. PTA-11344. The seeds depositedwith the ATCC on Sep. 20, 2010 were taken from a repository maintainedby Phytogen Seed Company since before the filing date of thisapplication. Access to the ATCC deposit will be available during thependency of the application to the Commissioner of Patents andTrademarks and persons determined by the Commissioner to be entitledthereto upon request. Upon allowance of any claims in the application,the Applicant(s) will maintain and will make this deposit available tothe public pursuant to the Budapest Treaty.

III. Processes of Preparing Novel Cotton Plants

-   -   A. Novel Cotton Plants Obtained From Variety P07X.8212.RF

Various breeding schemes may be used to produce new cotton varietiesfrom cotton variety P07X.8212.RF. In one method, generally referred toas the pedigree method, P07X.8212.RF may be crossed with anotherdifferent cotton plant such as a second parent cotton plant variety,which either itself exhibits one or more selected desirablecharacteristic(s) or imparts selected desirable characteristic(s) to ahybrid combination. Examples of potentially desired characteristicsinclude greater yield, better stalks, better roots, reduced time to cropmaturity, better fiber quality (e.g. fineness, length, lengthuniformity, strength, reflectance), better storm resistance, betteragronomic quality, higher nutritional value, improved oil profile,reduced gossypol levels, resistance and/or tolerance to insecticides,herbicides, pests, heat and drought, and disease, and uniformity ingermination times, stand establishment, growth rate, maturity and bollsize. If the two original parent cotton plants do not provide all thedesired characteristics, then other sources can be included in thebreeding population. Elite varieties can also be used as startingmaterials for breeding or source populations from which to develop newvarieties.

Thereafter, resulting seed is harvested and resulting superior progenyplants are selected and selfed or sib-mated in succeeding generations,such as for about 5 to about 7 or more generations, until a generationis produced that no longer segregates for substantially all factors forwhich the varietal parents differ, thereby providing a large number ofdistinct, pure-breeding varieties.

In another embodiment for generating new cotton varieties, generallyreferred to as backcrossing, one or more desired traits may beintroduced into parent cotton plant variety P07X.8212.RF (the recurrentparent) by crossing the P07X.8212.RF plants with another cotton plant(referred to as the donor or non-recurrent parent), which carries thegene(s) encoding the particular trait(s) of interest to produce F₁progeny plants. Both dominant and recessive alleles may be transferredby backcrossing. The donor plant may also be a varietal cotton plant,but in the broadest sense can be a member of any plant variety orpopulation cross-fertile with the recurrent parent. Next, F₁ progenyplants that have the desired trait are selected. Then, the selectedprogeny plants are crossed with P07X.8212.RF to produce backcrossprogeny plants. Thereafter, backcross progeny plants comprising thedesired trait and the physiological and morphological characteristics ofcotton variety P07X.8212.RF are selected. This cycle is repeated forabout one to about eight cycles, preferably for about 3 or more times insuccession to produce selected higher backcross progeny plants thatcomprise the desired trait and all of the physiological andmorphological characteristics of cotton variety P07X.8212.RF listed inTable 1 as determined at the 5% significance level when grown in thesame environmental conditions. Exemplary desired trait(s) include insectresistance, cytoplasmic male sterility, enhanced fiber quality, enhancednutritional quality, herbicide resistance, yield stability, yieldenhancement, storm resistance, and resistance to bacterial, fungal,nematode and viral disease. One of ordinary skill in the art of plantbreeding would appreciate that a breeder uses various methods to helpdetermine which cotton plants should be selected from the segregatingpopulations and ultimately which varieties will be used commercially andwill be used to develop hybrids for commercialization. In addition tothe knowledge of the germplasm and other skills the breeder uses, a partof the selection process is dependent on experimental design coupledwith the use of statistical analysis. Experimental design andstatistical analysis are used to help determine which plants, whichfamily of plants, and finally which varieties and hybrid combinationsare significantly better or different for one or more traits ofinterest. Experimental design methods are used to assess error so thatdifferences between two varieties or two hybrid lines can be moreaccurately determined. Statistical analysis includes the calculation ofmean values, determination of the statistical significance of thesources of variation, and the calculation of the appropriate variancecomponents. Either a five or a one percent significance level iscustomarily used to determine whether a difference that occurs for agiven trait is real or due to the environment or experimental error. Oneof ordinary skill in the art of plant breeding would know how toevaluate the traits of two plant varieties to determine if there is nosignificant difference between the two traits expressed by thosevarieties. For example, see Fehr, Walt, Principles of CultivarDevelopment, p. 261-286 (1987), which is incorporated herein byreference in its entirety. Mean trait values may be used to determinewhether trait differences are significant, and preferably the traits aremeasured on plants grown under the same environmental conditions.

This method results in the generation of cotton plants withsubstantially all of the desired morphological and physiologicalcharacteristics of the recurrent parent and the particular transferredtrait(s) of interest. Because such cotton plants are heterozygous forloci controlling the transferred trait(s) of interest, the lastbackcross generation would subsequently be selfed to provide purebreeding progeny for the transferred trait(s).

Backcrossing may be accelerated by the use of genetic markers such asSSR, RFLP, SNP or AFLP markers to identify plants with the greatestgenetic complement from the recurrent parent.

Direct selection may be applied where a single locus acts as a dominanttrait, such as the herbicide resistance trait. For this selectionprocess, the progeny of the initial cross are sprayed with the herbicidebefore the backcrossing. The spraying eliminates any plants that do nothave the desired herbicide resistance characteristic, and only thoseplants that have the herbicide resistance gene are used in thesubsequent backcross. In the instance where the characteristic beingtransferred is a recessive allele, it may be necessary to introduce atest of the progeny to determine if the desired characteristic has beensuccessfully transferred. The process of selection, whether direct orindirect, is then repeated for all additional backcross generations.

It should be appreciated by those having ordinary skill in the art thatbackcrossing can be combined with pedigree breeding as where varietyP07X.8212.RF is crossed with another cotton plant, the resultant progenyare crossed back to variety P07X.8212.RF and thereafter, the resultingprogeny of this single backcross are subsequently inbred to develop newvarieties. This combination of backcrossing and pedigree breeding isuseful when recovery of fewer than all of the P07X.8212.RFcharacteristics than would be obtained by a conventional backcross isdesired.

In an additional embodiment of the present disclosure, new cottonvarieties can be developed by a method generally referred to as haploidbreeding. In this methodology, haploid plants are generated fromdiploid, heterozygous cotton plants that result from crossing cottonplant variety P07X.8212.RF with another, different cotton plant. Suchhaploid cotton plants may be generated by methods known to those skilledin the art such as by culturing haploid anthers or embryos from adiploid plant. Alternately, such haploid cotton plant may be generatedby crossing the diploid heterozygous cotton plant with a cotton plantthat comprises a haploid inducing gene, which, when present in thefemale parent results in offspring with a greatly enhanced frequency ofhaploids of both maternal and paternal origin. Thereafter, homozygousdiploid plants are produced by the doubling of a set of chromosomes (1N)from a haploid plant generated by self-pollination such as through useof a doubling agent, such as colchicine, nitrous oxide gas, heattreatment and trifluralin. The technique of haploid breeding isadvantageous because no subsequent inbreeding is required to obtain ahomozygous plant from a heterozygous source. Thus, in another aspect ofthis disclosure a new cotton plant variety is developed by a method thatincludes the steps of crossing P07X.8212.RF or a hybrid made withP07X.8212.RF with another cotton plant having a propensity to generatehaploids to produce haploid progeny plants, and selecting desirablecotton plants from the haploid progeny plants.

Embodiments of the present disclosure also relate to novel cotton plantsproduced by a method generally referred to as mutation breeding wherebyone or more new traits may be artificially introduced into cottonvariety P07X.8212.RF. The goal of artificial mutagenesis is to increasethe rate of mutation for a desired characteristic. Mutation rates can beincreased by many different means including temperature, long-term seedstorage, tissue culture conditions, radiation; such as X-rays, Gammarays (e.g. Cobalt-60 or Cesium-137), neutrons, (product of nuclearfission by Uranium-235 in an atomic reactor), Beta radiation (emittedfrom radioisotopes such as Phosphorus-32 or Carbon-14), or ultravioletradiation (preferably from 2500 to 2900 nm), or chemical mutagens (suchas base analogues (5-bromo-uracil), related compounds (8-ethoxycaffeine), antibiotics (streptonigrin), alkylating agents (sulfurmustards, nitrogen mustards, epoxides, ethylenamines, sulfates,sulfonates, sulfones, lactones), azide, hydroxylamine, nitrous acid, oracridines. Once a desired trait is observed through mutagenesis andselected, the trait may then be incorporated into existing germplasm bytraditional breeding techniques. Details of mutation breeding can befound in “Principles of Cultivar Development”, Fehr, 1993 MacmillanPublishing Company, the disclosure of which is incorporated herein byreference in its entirety.

The mutagenesis treatment may be applied to various stages of plantdevelopment, including but not limited to cell cultures, embryos,microspores and shoot apices as well as to cotton seeds. By way ofexample, pollen may be mixed with a solution of 1 ml EMS and 100 mlsFisher paraffin oil (stock diluted by 1 ml and 15 mls oil solution)every minute for the first 5 minutes and then every five minutes for 45minutes to keep the pollen suspended. Thereafter, the pollen/paraffinoil solution is brushed onto the stigmas of emasculated flower buds. Apaper soda straw is used to cover the stigma to prevent contamination.The cotton boll is picked at maturity and then resultant seeds or theplants therefrom are screened for the desired mutant trait(s).

Once new varieties are created; the next step is to determine if the newvarieties have any value. This is accomplished by techniques ofmeasuring the combining ability of the new varietal plant, as well asthe performance of the variety itself. Combining ability refers to avariety's contribution as a parent when crossed with other varieties toform hybrids. Specific combining ability (SCA) refers to the ability ofa variety to cross to another specific variety to form a hybrid. Generalcombining ability (GCA) refers to the ability of a variety to cross to awide range of varieties to form hybrids. The methodology of forminghybrids to evaluate a variety's contribution as a parent for the purposeof selecting superior varieties is interchangeably known asexperimental, top or test crossing.

-   -   B. Novel Varieties Obtained From A Hybrid Having Variety        P07X.8212.RF As A Parent

In accordance with embodiments of the present disclosure, a hybrid planthaving variety P07X.8212.RF as a parent is crossed with itself or anydifferent cotton plant such as a varietal cotton plant or a hybridcotton plant to develop a novel variety. For example, a hybrid cottonplant having cotton plant variety P07X.8212.RF as a parent may beinbred, i.e., crossed to itself or sib-pollinated, and the resultingprogeny each selfed for about 5 to about 7 or more generations, therebyproviding a set of distinct, relatively pure-breeding varieties whereineach of the varieties received all of its alleles from the hybrid cottonplant having cotton plant variety P07X.8212.RF as a parent. Doublehaploid methods can also be used to obtain a cotton plant variety thatis homozygous at essentially every locus, wherein the cotton plantvariety received all of its alleles from the hybrid cotton plant havingcotton plant P07X.8212.RF as a parent. In other embodiments, a hybridcotton plant having cotton plant variety P07X.8212.RF as a parent iscrossed with a different cotton plant that may include any varietalcotton plant that is not varietal cotton plant P07X.8212.RF, any hybridcotton plant that does not have P07X.8212.RF as a parent, anothergermplasm source, a haploid or mutation inducing stock, or a trait donorplant, thereby providing a set of distinct, relatively pure-breedingvarieties. The resulting varieties can then be crossed with othervarieties or other cotton germplasm and the resulting progeny analyzedfor beneficial characteristics. In this way, novel varieties conferringdesirable characteristics can be identified.

-   -   C. “Chasing Selfs”

In the event that commercial cotton hybrids are developed, both femaleand male varietal seed may occasionally be found within a commercial bagof hybrid seed. Chasing the selfs involves identifying parental varietalplants within a stand of cotton that has been grown from a bag of hybridcotton seed. Once the seed is planted, the parental plants may beidentified and selected due to their variance from the population norm,i.e., by their stature, fruiting branch structure, leaf shape, leafpubescence, fiber quality traits, or yield components relative to thehybrid plants that grow from the hybrid seed that predominates in acommercial bag of hybrid seed. By locating the parental plants,isolating them from the rest of the plants, and self-pollinating them(i.e., “chasing selfs”), a breeder can obtain a variety that isidentical to a parent used to produce the hybrid.

Accordingly, another embodiment of the present disclosure is directed toa method for producing cotton plant variety P07X.8212.RF comprising: (a)planting a collection of seed, such as a collection of seed comprisingseed of a hybrid, one of whose parents is cotton variety P07X.8212.RF,the collection also comprising seed of the variety; (b) growing plantsfrom said collection of seed; (c) identifying parent plants; (d)controlling pollination in a manner that preserves substantialhomozygosity of the parent plant; and, (e) harvesting resultant seed.Step (c) may further comprise identifying plants with decreased vigor,i.e., plants that appear less robust than the other plants, oridentifying plants that have a genetic profile in accordance with thegenetic profile of P07X.8212.RF. Cotton plants capable of expressingsubstantially all of the physiological and morphological characteristicsof cotton variety P07X.8212.RF include cotton plants obtained by chasingselfs from a bag of hybrid seed.

One having skill in the art will recognize that once a breeder hasobtained cotton variety P07X.8212.RF by chasing selfs from a bag ofhybrid seed, the breeder can then produce new varietal plants such as bysib-pollinating, i.e., crossing the cotton plant P07X.8212.RF withanother cotton plant P07X.8212.RF, or by crossing the cotton plantP07X.8212.RF with a hybrid cotton plant obtained by growing thecollection of seed.

IV. Novel Hybrid Plants

-   -   A. Novel Hybrid Seeds and Plants

In yet another aspect of the disclosure, processes are provided forproducing cotton seeds or plants, which processes generally comprisecrossing a first parent cotton plant with a second parent cotton plantwherein at least one of the first parent cotton plant or the secondparent cotton plant is parent cotton plant variety P07X.8212.RF. In someembodiments of the present disclosure, the first cotton plant variety isP07X.8212.RF and is a female and in other embodiments the first cottonplant variety is P07X.8212.RF and is a male. These processes may befurther exemplified as processes for preparing hybrid cotton seed orplants, wherein a first cotton plant variety is crossed with a secondcotton plant of a different, distinct variety to provide a hybrid thathas, as one of its parents, the cotton plant variety P07X.8212.RF. Inthis case, a second variety is selected that confers desirablecharacteristics when in hybrid combination with the first variety. Inthese processes, crossing will result in the production of seed andlint. The seed and lint production occurs regardless whether the seedand/or lint is collected.

Any time the cotton plant variety P07X.8212.RF is crossed with another,different cotton variety, a first generation (F₁) cotton hybrid plant isproduced. As such, an F₁ hybrid cotton plant may be produced by crossingP07X.8212.RF with any second cotton plant variety. Therefore, any F₁hybrid cotton plant or cotton seed that is produced with P07X.8212.RF asa parent is part of the present disclosure.

When cotton plant variety P07X.8212.RF is crossed with another cottonplant variety to yield a hybrid, the original variety can serve aseither the maternal or paternal plant with basically, the samecharacteristics in the hybrids. Occasionally, maternally inheritedcharacteristics may express differently depending on the decision ofwhich parent to use as the female. However, often one of the parentalplants is preferred as the maternal plant because of increased seedand/or lint yield and preferred production characteristics, such asoptimal seed size and quality or ease of boll or lint removal.Particularly in very hot climates, such as in the Southwest USA, pollencan be shed better by one plant, rendering that plant the preferred maleparent. It is generally preferable to use P07X.8212.RF as the maleparent.

In embodiments of the present disclosure, the first step of “crossing”the first and the second parent cotton plants comprises planting,preferably in pollinating proximity, seeds of a first cotton plantvariety and a second, distinct cotton plant variety. As discussedherein, the seeds of the first cotton plant variety and/or the secondcotton plant variety can be treated with compositions that render theseeds and seedlings grown therefrom more hardy when exposed to adverseconditions.

A further step comprises cultivating or growing the seeds of the firstand second parent cotton plants into plants that bear flowers. If theparental plants differ in timing of sexual maturity, techniques may beemployed to obtain an appropriate nick, i.e., to ensure the availabilityof pollen from the parent cotton plant designated the male during thetime at which stigmas on the parent cotton plant designated the femaleare receptive to the pollen. Methods that may be employed to obtain thedesired nick include delaying the flowering of the faster maturingplant, such as, but not limited to delaying the planting of the fastermaturing seed, cutting or burning the top leaves of the faster maturingplant (without killing the plant) or speeding up the flowering of theslower maturing plant, such as by covering the slower maturing plantwith film designed to speed germination and growth.

In a preferred embodiment, the cotton plants are treated with one ormore agricultural chemicals as considered appropriate by the grower.

A subsequent step comprises preventing self-pollination orsib-pollination of the plants, i.e., preventing the stigmas of a plantfrom being fertilized by any plant of the same variety, including thesame plant. This is preferably done in large scale production bycontrolling the male fertility, e.g., treating the flowers so as toprevent pollen production or alternatively, using as the female parent amale sterile plant of the first or second parent cotton plant (i.e.,treating or manipulating the flowers so as to prevent pollen production,to produce an emasculated parent cotton plant or using as a female, acytoplasmic male sterile version of the cotton plant). This control mayalso be accomplished in small scale production by physical removal ofthe staminal column of individual flowers before anthesis to provideeffective control of unwanted self-pollination or sib-pollination.

Yet another step comprises allowing cross-pollination to occur betweenthe first and second parent cotton plants. When the plants are not inpollinating proximity, this is done by either collecting ripe,undehisced anthers from a flower on the pollen parent with a shortsection of a soda straw during the same evening of the emasculations, orcollecting whole, freshly dehisced flowers during the next morning afterthe emasculations. The soda straw containing the ripe anthers is thenslipped over the stigma of an emasculated flower. Finally, bracts arewired around the soda straw, holding it in place over the style, thusprotecting the stigma from foreign pollen. If a whole flower from themale parent is used, the petals are folded down and the staminal columnis rubbed onto the emasculated stigma. In small scale production, seedsof hybrid cotton are commercially produced by hand emasculation andpollination, or by hand pollination of genetic male-sterile cotton. Inlarge scale production, seed of hybrid cotton are commercially producedby using various bee and other insect pollinators to cross pollinategenetic or cytoplasmic male-sterile cotton, or cotton that has beentreated with a chemical that results in male sterility.

A further process comprises harvesting the seeds and/or lint, near or atmaturity, from the bolls of the plants that received the pollen. In aparticular embodiment, seed and/or lint is harvested from the femaleparent plant, and when desired, the harvested seed can be grown toproduce a first generation (F₁) hybrid cotton plant.

Yet another process comprises ginning the seed cotton to separate theseed from the marketable lint and delinting the “fuzzy” seed to removethe short “linters” that remain attached after ginning. The seeds arefurther conditioned and treated with chemicals such as fungicides andinsecticides prior to being packaged for sale to growers for theproduction of lint and seed. As with varietal seed, it may be desirableto treat hybrid seeds with compositions that render the seeds andseedlings grown therefrom more hardy when exposed to adverse conditions.The resulting hybrid seed is sold to growers for the production of seedand lint and not generally for breeding.

Still further, embodiments of the present disclosure provide a hybridcotton plant produced by growing the harvested seeds produced on themale-sterile plant, as well as seed produced by the hybrid cotton plant.

A single cross hybrid is produced when two different parent cotton plantvarieties are crossed to produce first generation F₁ hybrid progeny.Generally, each parent cotton plant variety has a genotype thatcomplements the genotype of the other parent variety. Typically, the F₁progeny are more vigorous than the respective parent cotton plantvarieties. This hybrid vigor, or heterosis, is manifested in manypolygenic traits, including markedly improved yields and improvedfruiting, roots, uniformity and insect and disease resistance. It is forthis reason that single cross F₁ hybrids are generally the most soughtafter hybrid. A three-way, or modified single-cross hybrid is producedfrom three varieties where two of the varieties are crossed (A×B) andthen the resulting F₁ hybrid is crossed with the third variety (A×B)×C,as where a modified female is used in the cross. A modified femaleprovides an advantage of improved seed/lint parent yield whereas amodified male improves pollen flow. A double cross hybrid is producedfrom four varieties crossed in pairs (A×B and C×D), thereby resulting intwo F₁ hybrids that are crossed again. Double cross hybrids are morecommon in countries wherein less demand exists for higher yieldingsingle cross hybrids. Synthetic populations or crosses are developed bycrossing two or more varieties (or hybrids, or germplasm sources)together and then employing one of many possible techniques to randommate the progeny. Random mating the progeny is any process used by plantbreeders to make a series of crosses that will create a new germplasmpool from which new breeding germplasm can be derived. Since crosspollination of male sterile cotton plants by hand or by various insectsis generally very inefficient, F₁ hybrid seed is generally too expensiveto produce on a large scale. Consequently, the F₂ seed harvested from F₁hybrids may retain suitable heterosis to be an economically viableoption to pure-line varieties.

The utility of the cotton plant variety P07X.8212.RF also extends tocrosses with species other than the barbadense species, such ashirsutum. Commonly, suitable species will be of the family Malvaceae,and especially of the genera Gossypium.

-   -   B. Cotton Varietal Comparison

As mentioned above, experimental strains are progressively eliminatedfollowing detailed evaluations of their phenotype, including formalcomparisons with other commercially successful varieties. Researchsmall-plot trials and commercial strip trials are used to compare thephenotypes of varieties grown in as many environments as possible. Theyare performed in many environments to assess overall performance of thenew varieties and to select optimum growing conditions. Because thecotton strains and varieties are grown in close proximity, differentialeffects of environmental factors that affect gene expression, such asmoisture, temperature, sunlight, and pests, are minimized For a decisionto be made to advance a strain, it is not necessary that the strain bebetter than all other varieties. Rather, significant improvements mustbe shown in at least some traits that would create value for someapplications or markets. Some experimental strains are eliminateddespite being similarly competitive relative to the current commercialvarieties because of the cost to bring a new variety to market requiresa new product to be a significant improvement over the existing productoffering. Such varieties may also be licensed to other parties who havea need in their commercial product portfolio.

P07X.8212.RF was evaluated in multiple over-years performance trials andcompared to PHY 800, the leading market variety and Pima yield standardfor the San Joaquin Valley, Calif. P07X.8212.RF had lint yieldcomparable to PHY 800 and good stability of performance over years(Table 2).

TABLE 2 Mean Lint Yield of P07X.8212.RF Compared to PHY 800 (Years2007-2009) Locations P07X.8212.RF PHY 800 P07X.8212.RF to (California)(lb/ac) (lb/ac) LSD (P = .05) CV (%) PHY 800 Ratio 2007 Buena Vista 26332771 221 6.1 95 Buttonwillow 1965 2059 259 9.9 95 Corcoran 1933 2028 1887.4 95 Stratford 2828 2601 345 10.6 109 Over Locations 2340 2365 205 8.699 2008 Buena Vista* 2581 2208 253 7.8 117 Kern Lake 1212 1085 177 10.9112 Corcoran ER 1815 1845 213 9.8 98 Corcoran PR 1760 1869 189 8.6 94Stratford 1734 1828 220 9.1 95 Over Locations 1820 1767 185 9.1 103 2009Buena Vista 1967 1904 178 6.7 103 Kern Lake 1805 1757 208 8.6 103Corcoran HM 2038 2119 240 8.1 96 Corcoran ER 1653 1487 184 8.9 111Corcoran PR 1327 1381 186 10.3 96 Corcoran TU 1307 1214 164 8.9 108 OverLocations 1683 1644 124 8.5 102 2007-2009 Overall 1904 1877 82 8.2 101*Significant difference at P = .05 Abbreviations: ac, acre; CV,coefficient of variation; lb, pound; LSD, least significant difference;P, probability of a greater difference

P07X.8212.RF has shown potential for earlier maturity (greaterpercentage first pick) compared to PHY 800 in certain growingenvironments (Table 3).

TABLE 3 First Pick (%) of P07X.8212.RF Compared to PHY 800 in 2008P07X.8212.RF Locations P07X.8212.RF PHY 800 LSD to PHY 800 (California)(%) (%) (P = .05) CV (%) Ratio Corcoran - El Rico* 92.70 89.86 1.38 0.8103 Corcoran - Stevenson 96.11 96.27 1.24 0.7 100 Corcoran - Paso Robles92.43 92.37 0.95 0.5 100 Corcoran - Tulare* 87.30 83.86 2.27 1.4 104Corcoran - Homeland 96.14 95.70 0.99 0.6 100 Over Locations (2008)*92.94 91.61 0.57 0.8 101 *Significant difference at P = .05Abbreviations: CV, coefficient of variation; LSD, least significantdifference; P, probability of a greater difference

P07X.8212.RF has similar boll weight and seed index, and significantlyhigher lint percentage and gin turn-out compared to PHY 800 (Table 4).

TABLE 4 Boll and Seed Traits of P07X.8212.RF Compared to PHY 800 in 2008Trait P07X.8212.RF PHY 800 LSD (P = .05) CV (%) Boll Weight (g) 3.753.77 0.17 10.0 Seed Index 13.7 13.8 0.3 4.2 (g/100 seeds) Lint (%)* 39.939.0 0.3 1.4 Gin Turn-Out* 32.5 31.5 0.8 2.2 *Significant difference atP = .05 Abbreviations: CV, coefficient of variation; LSD, leastsignificant difference; P, probability of a greater difference

The lint produced from P07X.8212.RF has improved fiber qualitiescompared to PHY 800. P07X.8212.RF has significantly longer fiber length,higher uniformity, higher fiber strength, higher elongation, and lessyellowness than PHY 800 (Table 5). These are important traits in thetextile cotton industry and are highly desired by market end-users. Lintcontaining this combination of fiber properties has heretofore not beenreported in any commercial cotton variety.

TABLE 5 Fiber Traits of P07X.8212.RF Compared to PHY 800 Over Locationsand Years (2007-2008) LSD CV Fiber Traits P07X.8212.RF PHY 800 (P = .05)(%) 2007 HVI Measurements, Samples From 4 Field Locations Fiber length(inch)* 1.44 1.41 0.01 1.4 Uniformity (%)* 88.8 88.1 0.6 1.0 Strength(g/Tex)* 45.3 43.1 1.2 3.9 Elongation (% change) 6.04 5.79 0.26 6.2Micronaire 4.08 4.11 0.09 3.1 Reflectance (% Rd) 72.8 72.5 0.7 1.4Yellowness (Hunter's + b)*^(a) 11.4 11.6 0.2 2.4 2008 HVI Measurements,Samples From 5 Field Locations Fiber length (inch)* 1.46 1.43 0.01 1.4Uniformity (%)* 89.2 88.5 0.5 0.9 Strength (g/Tex)* 48.2 46.8 1.2 4.1Elongation (% change)* 6.22 5.98 0.24 6.4 Micronaire 4.10 4.08 0.10 3.9Reflectance (% Rd) 73.5 73.1 0.6 1.4 Yellowness (Hunter's + b)^(a) 11.211.3 0.2 2.5 Overall 2007-2008 HVI Measurements Fiber length (inch)*1.45 1.42 0.01 1.4 Uniformity (%)* 89.0 88.3 0.4 1.0 Strength (g/Tex)*46.9 45.2 0.9 4.0 Elongation (% change)* 6.14 5.89 0.17 6.4 Micronaire4.09 4.09 0.07 3.6 Reflectance (% Rd) 73.2 72.8 0.5 1.4 Yellowness(Hunter's + b)*^(a) 11.3 11.4 0.1 2.4 *Significant difference at P = .05Abbreviations: HVI, High Volume Instrument; LSD, least significantdifference; Rd, diffuse reflectance ^(a)Lower value is an improvement

Both P07X.8212.RF and PHY 800 are considered to have tolerance to thepathogen, Fusarium oxysporum vasinfectum, Race 4 (FOV Race 4). Data fromFOV Race-4 field and greenhouse screenings demonstrate P07X.8212.RF andPHY 800 have significantly less foliar and root damage and significantlygreater field-stand survival than the susceptible variety DPL 744 (Table6).

TABLE 6 Fusarium oxysporum vasinfectum Race-4 Screens for P07X.8212.RFand PHY 800 Compared to Susceptible Variety DPL 744 LSD ToleranceMeasure P07X.8212.RF PHY 800 DPL 744 (P = .05) Fresno County FieldScreen Foliar Damage Index  0.42*  0.00* 3.15 1.16 Root Vascular  0.88* 0.40* 3.87 0.71 Stain Index Stand Count 93.3*  97.1*  48.4 14.6Survival (%) Kearney Greenhouse Screen Foliar Damage Index  0.22*  0.59*2.86 0.65 Root Vascular  0.61*  0.95* 3.55 0.72 Stain Index*Significantly different from DPL 744 at P = .05

P07X.8212.RF is similar to the commercial variety PHY 800 in growthhabit and structure. P07X.8212.RF tends to have a slightly shorteraverage plant height, less total nodes and vegetative nodes, more bollsper plant, more first position bolls, yet a lower percentage firstposition bolls in comparison to PHY 800; though not statisticallydifferent on any measured parameter (Table 7).

TABLE 7 Morphology of P07X.8212.RF Compared to PHY 800 Averaged Over 2Locations near Corcoran, California LSD CV Morphological TraitP07X.8212.RF PHY 800 (P = .05) (%) Plant Height (cm) 89.2 94.2 7.5 9.1Total Nodes 20.1 20.4 1.3 6.9 No. Vegetative Nodes 6.3 6.7 0.7 12.6 No.Fruiting Nodes 13.8 13.7 1.4 10.7 Mean Internode 4.4 4.6 0.3 6.6 Length(cm) Bolls per plant 20.7 18.0 3.4 19.9 No. First Position Bolls 11.310.7 1.3 13.6 % First Position Bolls* 55.2 61.5 6.3 12.1 Locules PerBoll 3.00 3.02 0.04 2.2 *Significant difference at P = .05Abbreviations: cm, centimeter; CV, coefficient of variation; LSD, leastsignificant difference; No., number of; P, probability of a greaterdifference

P07X.8212.RF appears stable and uniform after four generations ofisolated field seed production and field-trial evaluations, and nooff-type plants have been exhibited. This line has exhibited commercialvalue in multi-year and multi-location field evaluations.

V. Novel P07X.8212.RF-Derived Plants

All plants produced using cotton plant variety P07X.8212.RF as a parentare within the scope of embodiments of this disclosure, including plantsderived from cotton plant variety P07X.8212.RF. This includes plantsessentially derived from variety P07X.8212.RF with the term “essentiallyderived variety” having the meaning ascribed to such term in 7 U.S.C.§2104(a)(3) of the Plant Variety Protection Act, which section is herebyincorporated by reference in its entirety. This also includes progenyplants and parts thereof with at least one ancestor that is cotton plantvariety P07X.8212.RF and more specifically where the pedigree of thisprogeny includes 1, 2, 3, 4, and/or 5 or cross pollinations to cottonplant P07X.8212.RF, or a plant that has P07X.8212.RF as a progenitor.All breeders of ordinary skill in the art maintain pedigree records oftheir breeding programs. These pedigree records contain a detaileddescription of the breeding process, including a listing of all parentallines used in the breeding process and information on how such line wasused. Thus, a breeder would know if P07X.8212.RF were used in thedevelopment of a progeny line, and would also know how many breedingcrosses to a line other than P07X.8212.RF were made in the developmentof any progeny line. A progeny line so developed may then be used incrosses with other, different, cotton varieties to produce firstgeneration F1 cotton hybrid seeds and plants with superiorcharacteristics.

Accordingly, another aspect of the present disclosure is methods forproducing a P07X.8212.RF-derived cotton plant. Embodiments of suchmethods for producing a P07X.8212.RF-derived cotton plant, comprise: (a)crossing cotton plant P07X.8212.RF with a second cotton plant to yieldprogeny cotton seed; and (b) growing the progeny cotton seed, (underplant growth conditions), to yield the P07X.8212.RF-derived cottonplant. Such methods may further comprise the steps of: (c) crossing theP07X.8212.RF-derived cotton plant with itself or another cotton plant toyield additional P07X.8212.RF-derived progeny cotton seed; (d) growingthe progeny cotton seed of step (b) (under plant growing conditions), toyield additional P07X.8212.RF-derived cotton plants; and (e) repeatingthe crossing and growing steps of (c) and (d) from 0 to 7 times togenerate further P07X.8212.RF-derived cotton plants. Still further, thismay comprise utilizing methods of semigamy and other haploid breedingand plant tissue culture methods to derive progeny of theP07X.8212.RF-derived cotton plant.

VI. Tissue Cultures and In Vitro Regeneration of Cotton Plants

As is well known in this art, tissue culture of cotton may be used forthe in vitro regeneration of a cotton plant. Accordingly, a furtheraspect of the disclosure relates to tissue cultures of the cotton plantvariety designated P07X.8212.RF, to tissue cultures of hybrid andderived cotton plants obtained from P07X.8212.RF, to plants obtainedfrom such tissue cultures and to the use of tissue culture methodologyin plant breeding. The term “tissue culture” includes a compositioncomprising isolated cells of the same type, isolated cells of adifferent type, or a collection of such cells organized into parts of aplant. Exemplary tissue cultures are protoplasts, calli and plant cellsthat are intact in plants or parts of plants, such as embryos, pollen,ovules, flowers, petals, seeds, bolls, gossypol glands, stems, leaves,fibers, roots, root tips, and the like. In a preferred embodiment, thetissue culture comprises embryos, protoplasts, meristematic cells,pollen, leaves or anthers derived from immature tissues of these plantparts.

-   -   A. Cotyledon Culture

To obtain plant tissue for callus culture initiation, seeds areharvested from a wild type cotton plant (generally GC510 or Coker310genotype). Initially, seeds are surface sterilized by a triple rinsewith 70% ethanol for 1 minute each, a thorough rinse with sterile water,followed by a wash in 30% commercial bleach (0.1% sodium hypochlorite)for about 20 minutes.

Seeds are rinsed in sterile distilled water, and seeds are placed on thesurface of germination media (LS salts (10×), 3% sucrose, modified B5vitamins (1000×), at pH 5.8) for the production of sterile plantlets.Approximately, 7-10 days post plating plantlets will have emerged fromthe seeds. The “first true leaves” are the cotyledons. Generally, tissueculture media contains amino acids, salts, sugars, hormones, andvitamins. The proportion of one ingredient versus another depends on theapplication (e.g., need for rooting versus shoot elongation). At day7-10, the cotyledons are of sufficient size for experiment use. Thecotyledons are cut into 1 mm square pieces and plated on callusinduction media (100 ml/L LS salts (10×), 3% glucose, 1 ml/L modified B5vitamins (1000×), 1 ml/L 1 mM kinetin, 1 ml/L 1 mM 2,4-D, 8 g/L nobleagar, pH 5.8). The cotyledon segment is placed on the media in theabaxial side down orientation. After three weeks on the callus inductionmedia, callus forms around the cut edges of the segment; the callus isremoved from the edges using a scalpel. The “callus” is a loosecollection or mass of undifferentiated cells, which can be yellow-greenin color. Some lines are prone to phenolic production (browning), whichcan affect growth. The callus is maintained on the initiation media fornine weeks, with subculture to fresh media every three weeks. If thesegments are treated with Agrobacterium, the callus induction mediainclude carbenicillin, an antibiotic to kill the Agrobacterium (2 ml/L),and glufosinate-ammonium (0.5 ml/L), which is the selective agent thatallows growth of only those cells that contain a transgene (PAT).

At week nine, the callus is transferred to a growth media (100 ml/L LSsalts, 3% glucose, 1 ml/L B5 vitamins, 4.6 ml/L kinetin, 10.7 ml/L NAA,8 g/L noble agar, pH 5.8) and, if Agrobacterium infection was used totransfer the PAT gene, carbenicillin (0.4 ml/L) and glufosinate ammonium(0.3 ml/L)). The callus should remain on this media for 3 weeks, toallow for increased growth before going to embryogenic callus inductionmedia. Once sufficient callus is present, the tissue is placed onembryogenic induction media (1 pkg DKW salts, 10 ml/L myo-inositol, 1ml/L B5 vitamins, 2% glucose, 8 g/L noble agar, pH 5.8). The time for aline to produce embryogenic callus varies from two to six months; duringthis time the callus remains on the same plate of media. Stress canassist in inducing cotton callus to become embryogenic.

Regeneration begins with embryogenic callus. Embryogenic callus ismaintained on the embryogenic callus induction media with two weeksubcultures to fresh media. Microscope use is preferred for theisolation and transfer of embryogenic callus to ensure the desiredmorphology is taken from the plates. The desired morphology has agranular appearance, yellow-green in color. The embryogenic callus willgive rise to embryos, which may look like small footballs and have moreof a green color. The embryos mature on the embryogenic callus inductionmedia. It may take three to nine weeks for the embryos to mature orelongate; transfers are carried out at three week intervals. At thisstage the embryos are transferred to a basal media that will improveshoot (1 pkg DKW salts, 10 ml/L myo-inositol, 1 ml/L modified B5vitamins, 3% sucrose, 0.5 ml/L kinetin, 8 g/L noble agar, pH 5.8) orroot development (0.5 pkg DKW salts, 5 m/L myo-inositol, 0.5 ml/Lmodified B5 vitamins, 1% sucrose, 8 g/L noble agar, pH5.8).

When secondary roots have formed and the shoot is 1 to 2 inches highwith 2 good leaves, the cotton plant is ready for soil. Plantlets arefirst placed in a Conviron in small pots with a humidi-dome to assistwith plant hardening, since cotton plants can be quite fragile. Thenplants are later transferred to large pots in the greenhouse. Mostcotton plants are allowed to self-pollinate and these flowers are taggedwith one color, while others may be crossed with an elite variety andtagged separately.

-   -   B. Additional Tissue Cultures and Regeneration

Other means for preparing and maintaining plant tissue cultures are wellknown in the art. By way of example, reference may be had to Komatsuda,T. et al., Crop Sci. 31:333-337 (1991); Stephens, P. A., et al., Theor.Appl. Genet. 82:633-635 (1991); Komatsuda, T. et al., Plant Cell, Tissueand Organ Culture, 28:103-113 (1992); Dhir, S. et al., Plant CellReports 11:285-289 (1992); Pandey, P. et al., Japan J. Breed. 42:1-5(1992); and Shetty, K., et al., Plant Science 81:245-251 (1992); as wellas U.S. Pat. No. 5,024,944 issued Jun. 18, 1991 to Collins et al., andU.S. Pat. No. 5,008,200 issued Apr. 16, 1991 to Ranch et al. Thus,another aspect of this disclosure is to provide cells that upon growthand differentiation produce cotton plants having the physiological andmorphological characteristics of the present cotton variety.

VII. Male Sterility

Methods for controlling male fertility in cotton plants offer theopportunity for improved plant breeding, particularly for thedevelopment of cotton hybrids that require the implementation of a malesterility system to prevent the varietal parent plants fromself-pollination.

Accordingly, another aspect of the present disclosure is male-sterilevarietal cotton plants designated P07X.8212.RF and the production ofhybrid cotton seed using a male sterility system with such varietalfemale parent plants that are male sterile. In the event that cottonvariety P07X.8212.RF is employed as the female parent, P07X.8212.RF canbe rendered male-sterile by, for example, removing the stamens ofP07X.8212.RF parental plants manually. By way of example, alternatestrips of two cotton varieties may be planted in a field followed bymanual emasculation. Provided that the female variety is sufficientlyisolated from foreign cotton pollen sources, the stigma of theemasculated variety will be fertilized only from the other male varietyeither manually or by insect pollinator vectors, and the resulting seedwill therefore be hybrid seed.

The laborious and occasionally unreliable manual emasculation processcan be minimized by using cytoplasmic male-sterile (CMS) varieties.Plants of a CMS variety are male sterile as a result of factorsresulting from cytoplasmic as opposed to the nuclear genome. Thus, thischaracteristic is inherited exclusively through the female parent incotton plants since CMS plants are fertilized with pollen from anothervariety that is not male-sterile. Pollen from the second variety may ormay not contribute genes that make the hybrid plants male-fertile. Seedfrom emasculated fertile cotton and CMS produced seed of the same hybridcan be blended to insure that adequate pollen loads are available forfertilization when the hybrid plants are grown. Conventionalbackcrossing methodology may be used to introgress the CMS trait intovariety P07X.8212.RF.

Alternatively, haploid breeding methods may also be employed to convertvariety P07X.8212.RF to CMS sterility. Haploids are plants that containonly one-half of the chromosome number present in diploid somatic cells,which are cells other than haploid cells, such as those found in thegerm. There are a few stocks or genetic systems in cotton that are knownto generate haploids spontaneously.

Manual emasculation can also be avoided by the use of chemically inducedmale sterility in the production of hybrid cotton seed. Chemicals thatinduce male sterility include gametocides, pollen suppressants, andchemical hybridizing agents. The general procedure is to use a foliarspray before flowering, which inhibits production of viable pollen, butdoes not injure the pistillate reproductive organs or affect seeddevelopment. If the treatment is successful and all of the pollenkilled, self-pollination will not occur in the treated plants, but theflowers will set seed freely from cross-pollination. In such a case, theparent plants used as the male may either not be treated with thechemical agent or may include a genetic factor that causes resistance tothe sterilizing effects of the chemical agent. The use of chemicallyinduced male sterility affects fertility in the plants only for thegrowing season in which the gametocide is applied.

The presence of a male-fertility restorer gene results in the productionof a fully fertile F₁ hybrid progeny. If no restorer gene is present inthe male parent, male-sterile hybrids are obtained. Such hybrids areuseful where the vegetative tissue of the cotton plant is used, e.g.,for silage, but in most cases, the seeds will be deemed the mostvaluable portion of the crop, so fertility of the hybrids in these cropsmust be restored. Therefore, one aspect of the present disclosureconcerns cotton variety P07X.8212.RF comprising a single gene capable ofrestoring male fertility in an otherwise male-sterile variety or hybridplant. Examples of male-sterility genes and corresponding restorers thatcould be employed within the scope of embodiments of the disclosure arewell known to those of skill in the art of plant breeding and aredisclosed in, for example, U.S. Pat. Nos. 5,530,191, 5,689,041,5,741,684, and 5,684,242, the disclosures of which are each specificallyincorporated herein by reference in their entirety.

VIII. Cotton Transformation

With the advent of molecular biological techniques that have allowed theisolation and characterization of genes that encode specific proteinproducts, scientists in the field of plant biology developed a stronginterest in engineering the genome of plants to contain and to expressforeign genes, or additional, or modified versions of native orendogenous genes (perhaps driven by different promoters) to alter thetraits of a plant in a specific manner. Such foreign, additional and/ormodified genes are referred to herein collectively as “transgenes.” Thepresent disclosure, in particular embodiments, also relates totransformed versions of the claimed cotton variety P07X.8212.RFcontaining one or more transgenes.

Plant transformation involves the construction of an expression vectorthat will function in plant cells. Such a vector comprises DNAcomprising a gene under control of or operatively linked to a regulatoryelement. The expression vector may contain one or more such operablylinked gene/regulatory element combinations. The vector(s) may be in theform of a plasmid, and can be used, alone or in combination with otherplasmids, to provide transformed cotton plants, using transformationmethods as described below to incorporate transgenes into the geneticmaterial of the cotton plant(s).

-   -   A. Expression Vectors for Cotton Transformation/Marker Genes

Expression vectors include at least one genetic marker, operably linkedto a regulatory element that allows transformed cells containing themarker to be either recovered by negative selection, i.e., inhibitinggrowth of cells that do not contain the selectable marker gene, or bypositive selection, i.e., screening for the product encoded by thegenetic marker. Many commonly used selectable marker genes for planttransformation are well known in the transformation arts, and include,for example, genes that code for enzymes that metabolically detoxify aselective chemical agent that may be an antibiotic or a herbicide, orgenes that encode an altered target that is insensitive to theinhibitor. A few positive selection methods are also known in the art.One commonly used selectable marker gene for plant transformation is theneomycin phosphotransferase II (nptll) gene, isolated from a bacterialsource, which when placed under the control of plant regulatory signalsconfers resistance to kanamycin. Fraley et al., Proc. Natl. Acad. Sci.U.S.A. 80: 4803 (1983). Another commonly used selectable marker gene isthe hygromycin phosphotransferase gene that confers resistance to theantibiotic hygromycin. Vanden Elzen et al., Plant Mol. Biol. 5: 299(1985).

Additional selectable marker genes of bacterial origin that conferresistance to antibiotics include gentamycin acetyl transferase,streptomycin phosphotransferase, aminoglycoside-3′-adenyl transferaseand the bleomycin resistance determinant. Hayford et al., Plant Physiol.86: 1216 (1988), Jones et al., Mol. Gen. Genet. 210: 86 (1987), Svab etal., Plant Mol. Biol. 14: 197 (1990), Hille et al., Plant Mol. Biol. 7:171 (1986). Other selectable marker genes confer resistance toherbicides such as glyphosate, glufosinate or bromoxynil. Comai et al.,Nature 317: 741-744 (1985), Gordon-Kamm et al., Plant Cell 2: 603-618(1990) and Stalker et al., Science 242 : 419-423 (1988).

Other selectable marker genes for plant transformation are not ofbacterial origin. These genes include, for example, mouse dihydrofolatereductase, plant 5-enolpyruvylshikimate-3-phosphate synthase and plantacetolactate synthase. Eichholtz et al., Somatic Cell Mol. Genet. 13: 67(1987), Shah et al., Science 233: 478 (1986), Charest et al., Plant CellRep. 8: 643 (1990).

Another class of marker genes for plant transformation require screeningof presumptively transformed plant cells rather than direct geneticselection of transformed cells for resistance to a toxic substance suchas an antibiotic. These genes are particularly useful to quantify orvisualize the spatial pattern of expression of a gene in specifictissues and are frequently referred to as reporter genes because theycan be fused to a gene or gene regulatory sequence for the investigationof gene expression. Commonly used genes for screening presumptivelytransformed cells include β-glucuronidase (GUS), β-galactosidase, andluciferase. Jefferson, R. A., Plant Mol. Biol. Rep. 5: 387 (1987), Teeriet al., EMBO J. 8: 343 (1989), Koncz et al., Proc. Natl. Acad. Sci.U.S.A. 84: 131 (1987). Another approach to the identification of arelatively rare transformation events has been use of a gene thatencodes a dominant constitutive regulator of the Zea mays anthocyaninpigmentation pathway. Ludwig et al., Science 247: 449 (1990).

Recently, in vivo methods for visualizing GUS activity that do notrequire destruction of plant tissue have been made available. MolecularProbes Publication 2908, Imagene Green™, p. 1-4 (1983) and Naleway etal., J. Cell Biol. 115: 151a (1991). However, these in vivo methods forvisualizing GUS activity have not proven useful for recovery oftransformed cells because of low sensitivity, high fluorescentbackgrounds, and limitations associated with the use of luciferase genesas selectable markers.

More recently, a gene encoding Green Fluorescent Protein (GFP) has beenutilized as a marker for gene expression in prokaryotic and eukaryoticcells. Chalfie et al., Science 263: 802 (1994). GFP and mutants of GFPcan be used as screenable markers.

-   -   B. Promoters

Genes included in expression vectors must be driven by a nucleotidesequence comprising a regulatory element, for example, a promoter.Several types of promoters are now well known in the transformationarts, as are other regulatory elements that can be used alone or incombination with promoters.

As used herein “promoter” includes reference to a region of DNA upstreamfrom the start of transcription and involved in recognition and bindingof RNA polymerase and other proteins to initiate transcription. A “plantpromoter” is a promoter capable of initiating transcription in plantcells. Examples of promoters under developmental control includepromoters that preferentially initiate transcription in certain tissues,such as leaves, roots, seeds, fibers, xylem vessels, tracheids, orsclerenchyma. Such promoters are referred to as “tissue-preferred.”Promoters that initiate transcription only in certain tissues arereferred to as “tissue-specific.” A “cell type” specific promoterprimarily drives expression in certain cell types in one or more organs,for example, vascular cells in roots or leaves. An “inducible” promoteris a promoter that is under environmental control or is induced inresponse to chemical or hormonal stimuli. Examples of environmentalconditions that may effect transcription by inducible promoters includeanaerobic conditions or the presence of light. Examples of chemicalsthat induce expression include salicyclic acid and ABA. Tissue-specific,tissue-preferred, cell type specific, and inducible promoters constitutethe class of “non-constitutive” promoters. A “constitutive” promoter isa promoter that is active under most environmental conditions and in allcells.

-   -   -   a. Inducible Promoters

An inducible promoter is operably linked to a gene for expression incotton. Optionally, the inducible promoter is operably linked to anucleotide sequence encoding a signal sequence that is operably linkedto a gene for expression in cotton. With an inducible promoter the rateof transcription increases in response to an inducing agent. Anyinducible promoter can be used in embodiments of the instant disclosure.A particularly preferred inducible promoter is a promoter that respondsto an inducing agent to which plants do not normally respond. Anexemplary inducible promoter is the inducible promoter from a steroidhormone gene, the transcriptional activity of which is induced by aglucocorticosteroid hormone.

-   -   -   2. Constitutive Promoters

A constitutive promoter is operably linked to a gene for expression incotton or the constitutive promoter is operably linked to a nucleotidesequence encoding a signal sequence that is operably linked to a genefor expression in cotton. Many different constitutive promoters can beused in embodiments of the present disclosure. Exemplary constitutivepromoters include, but are not limited to, the promoters from plantviruses such as the 35S promoter from CaMV and the promoters from suchgenes as rice actin, maize ubiquitin, and corn H3 histone. Also, the ALSpromoter, a XbaI/NcoI fragment 5′ to the Brassica napus ALS3 structuralgene (or a nucleotide sequence that has substantial sequence similarityto the XbaI/NcoI fragment) represents a particularly useful constitutivepromoter.

-   -   -   3. Tissue-specific or Tissue-Preferred Promoters

A tissue-specific promoter is operably linked to a gene for expressionin cotton. Optionally, the tissue-specific promoter is operably linkedto a nucleotide sequence encoding a signal sequence that is operablylinked to a gene for expression in cotton. Plants transformed with agene of interest operably linked to a tissue-specific promoter producethe protein product of the transgene exclusively, or preferentially, ina specific tissue. Any tissue-specific or tissue-preferred promoter canbe utilized in embodiments of the instant disclosure. Exemplarytissue-specific or tissue-preferred promoters include, but are notlimited to, a seed-preferred promoter such as that from the phaseolingene; a leaf-specific and light-induced promoter such as that from cabor rubisco; an anther-specific promoter such as that from LAT52; apollen specific promoter such as that from Zm13 or amicrospore-preferred promoter such as that from apg.

-   -   C. Signal Sequences For Targeting Proteins to Subcellular        Compartments

Transport of protein produced by transgenes to a subcellular compartmentsuch as the chloroplast, vacuole, peroxisome, glyoxysome, cell wall ormitochondrion, or for secretion into the apoplast, is accomplished bymeans of operably linking the nucleotide sequence encoding a signalsequence to the 5′ and/or 3′ region of a gene encoding the protein ofinterest. Targeting sequences at the 5′ and/or 3′ end of the structuralgene may determine, during protein synthesis and processing, where theencoded protein is ultimately compartmentalized. The presence of asignal sequence directs a polypeptide to either an intracellularorganelle or subcellular compartment or for secretion to the apoplast.Use of any signal sequence known in the art is contemplated inembodiments of the present disclosure.

-   -   D. Foreign Protein Genes and Agronomic Genes

Using transgenic plants in accordance with embodiments of the presentdisclosure, a foreign protein can be produced in commercial quantities.Thus, techniques for the selection and propagation of transformedplants, which are well understood in the art, yield a plurality oftransgenic plants, which are harvested in a conventional manner, and aforeign protein then can be extracted from a tissue of interest or fromtotal biomass. Protein extraction from plant biomass can be accomplishedby known methods.

According to a preferred embodiment, the transgenic plant provided forcommercial production of foreign protein is cotton. In another preferredembodiment, the biomass of interest is seed. For the relatively smallnumber of transgenic plants that show higher levels of expression, agenetic map can be generated, primarily via conventional RestrictionFragment Length Polymorphisms (RFLP), Polymerase Chain Reaction (PCR)analysis, and Simple Sequence Repeats (SSR), in a manner that identifiesthe approximate chromosomal location of the integrated DNA molecule. Forexemplary methodologies in this regard, see Glick and Thompson, Methodsin Plant Molecular Biology and Biotechnology 269-284 (CRC Press, BocaRaton, 1993). Map information concerning chromosomal location is usefulfor proprietary protection of a subject transgenic plant. Ifunauthorized propagation is undertaken and crosses made with othergermplasm, the map of the integration region can be compared to similarmaps for suspect plants, to determine if the latter have a commonparentage with the subject plant. Map comparisons can involvehybridizations, RFLP, PCR, SSR and sequencing, all of which areconventional techniques.

Likewise, in accordance with embodiments of the present disclosure,agronomic genes can be expressed in transformed plants. Moreparticularly, plants can be genetically engineered to express variousphenotypes of agronomic interest. Exemplary genes implicated in thisregard include, but are not limited to:

-   -   -   a. Genes that Confer Resistance to Pests or Disease and that            Encode:            -   i. Plant disease resistance genes. Plant defenses are                often activated by specific interaction between the                product of a disease resistance gene (R) in the plant                and the product of a corresponding avirulence (Avr) gene                in the pathogen. A plant variety can be transformed with                a cloned resistance gene to engineer plants that are                resistant to specific pathogen strains. See, for                example, Jones et al., Science 266: 789 (1994) (cloning                of the tomato Cf-9 gene for resistance to Cladosporium                fulvum); Martin et al., Science 262: 1432 (1993) (tomato                Pto gene for resistance to Pseudomonas syringae pv.                tomato encodes a protein kinase); Mindrinos et al., Cell                78: 1089 (1994) (Arabidopsis RSP2 gene for resistance to                Pseudomonas syringae).            -   ii. A Bacillus thuringiensis protein, a derivative                thereof or a synthetic polypeptide modeled thereon. See,                for example, Geiser et al., Gene 48: 109 (1986), who                disclose the cloning and nucleotide sequence of a Bt                δ-endotoxin gene. Moreover, DNA molecules encoding                δ-endotoxin genes can be purchased from American Type                Culture Collection (Rockville, Md.), for example, under                ATCC Accession Nos. 40098, 67136, 31995 and 31998.            -   iii. A lectin. See, for example, the disclosure by Van                Damme et al., Plant Molec. Biol. 24: 25 (1994), who                disclose the nucleotide sequences of several Clivia                miniata mannose-binding lectin genes; and Yenofsky et al                (U.S. Pat. No. 6,710,228), who disclose cotton cells,                plants and seeds genetically engineered to express                insecticidal and fungicidal chitin binding proteins.            -   iv. A vitamin-binding protein such as avidin. See PCT                application US93/06487, the contents of which are hereby                incorporated by reference in their entirety. The                application teaches the use of avidin and avidin                homologues as larvicides against insect pests.            -   v. An enzyme inhibitor, for example, a protease                inhibitor or an amylase inhibitor. See, for example, Abe                et al., J. Biol. Chem. 262: 16793 (1987) (nucleotide                sequence of rice cysteine proteinase inhibitor), Huub et                al., Plant Molec. Biol. 21: 985 (1993) (nucleotide                sequence of cDNA encoding tobacco proteinase inhibitor                I), and Sumitani et al., Biosci. Biotech. Biochem. 57:                1243 (1993) (nucleotide sequence of Streptomyces                nitrosporeus α-amylase inhibitor).            -   vi. An insect-specific hormone or pheromone such as an                ecdysteroid and juvenile hormone, a variant thereof, a                mimetic based thereon, or an antagonist or agonist                thereof. See, for example, the disclosure by Hammock et                al., Nature 344: 458 (1990), of baculovirus expression                of cloned juvenile hormone esterase, an inactivator of                juvenile hormone.            -   vii. An insect-specific peptide or neuropeptide that,                upon expression, disrupts the physiology of the affected                pest. For example, see the disclosures of Regan, J.                Biol. Chem. 269: 9 (1994) (expression cloning yields DNA                coding for insect diuretic hormone receptor), and Pratt                et al., Biochem. Biophys. Res. Comm 163: 1243 (1989) (an                allostatin is identified in Diploptera puntata). See                also U.S. Pat. No. 5,266,317 to Tomalski et al., who                disclose genes encoding insect-specific, paralytic                neurotoxins.            -   viii. An insect-specific venom produced in nature by a                snake, a wasp, etc. For example, see Pang et al., Gene                116: 165 (1992), for disclosure of heterologous                expression in plants of a gene coding for a scorpion                insectotoxic peptide.            -   ix. An enzyme responsible for an hyperaccumulation of a                monoterpene, a sesquiterpene, a steroid, hydroxamic                acid, a phenylpropanoid derivative or another                non-protein molecule with insecticidal activity.            -   x. An enzyme involved in the modification, including the                post-translational modification, of a biologically                active molecule, for example, a glycolytic enzyme, a                proteolytic enzyme, a lipolytic enzyme, a nuclease, a                cyclase, a transaminase, an esterase, a hydrolase, a                phosphatase, a kinase, a phosphorylase, a polymerase, an                elastase, a chitinase, or a glucanase, whether natural                or synthetic. See PCT application WO 93/02197 in the                name of Scott et al., which discloses the nucleotide                sequence of a callase gene. DNA molecules that contain                chitinase-encoding sequences can be obtained, for                example, from the ATCC under Accession Nos. 39637                and 67152. See also Kramer et al., Insect Biochem.                Molec. Biol. 23: 691 (1993), who teach the nucleotide                sequence of a cDNA encoding tobacco hookworm chitinase,                and Kawalleck et al., Plant Molec. Biol. 21: 673 (1993),                who provide the nucleotide sequence of the parsley                ubi4-2 polyubiquitin gene.            -   xi. A molecule that stimulates signal transduction. For                example, see the disclosure by Botella et al., Plant                Molec. Biol. 24: 757 (1994), of nucleotide sequences for                mung bean calmodulin cDNA clones, and Griess et al.,                Plant Physiol. 104: 1467 (1994), who provide the                nucleotide sequence of a corn calmodulin cDNA clone.            -   xii. A hydrophobic moment peptide. See PCT application                WO95/16776 (disclosure of peptide derivatives of                Tachyplesin that inhibit fungal plant pathogens) and PCT                application WO95/18855 (teaches synthetic antimicrobial                peptides that confer disease resistance), the respective                contents of which are hereby incorporated by reference                in their entirety.            -   xiii. A membrane permease, a channel former or a channel                blocker. For example, see the disclosure by Jaynes et                al., Plant Sci. 89: 43 (1993), of heterologous                expression of a cecropin-β lytic peptide analog to                render transgenic tobacco plants resistant to                Pseudomonas solanacearum.            -   xiv. A viral-invasive protein or a complex toxin derived                therefrom. For example, the accumulation of viral coat                proteins in transformed plant cells imparts resistance                to viral infection and/or disease development effected                by the virus from which the coat protein gene is                derived, as well as by related viruses. See Beachy et                al., Ann. Rev. Phytopathol. 28: 451 (1990). Coat                protein-mediated resistance has been conferred upon                transformed plants against alfalfa mosaic virus,                cucumber mosaic virus, tobacco streak virus, potato                virus X, potato virus Y, tobacco etch virus, tobacco                rattle virus and tobacco mosaic virus. Id.            -   xv. An insect-specific antibody or an immunotoxin                derived therefrom. Thus, an antibody targeted to a                critical metabolic function in the insect gut would                inactivate an affected enzyme, killing the insect. Cf.                Taylor et al., Abstract #497, Seventh Intl. Symposium on                Molecular Plant-Microbe Interactions (Edinburgh,                Scotland (Edinburgh, Scotland, 1994) (enzymatic                inactivation in transgenic tobacco via production of                single-chain antibody fragments).            -   xvi. A virus-specific antibody. See, for example,                Tavladoraki et al, Nature 366: 469 (1993), who show that                transgenic plants expressing recombinant antibody genes                are protected from virus attack.            -   xvii. A developmental-arrestive protein produced in                nature by a pathogen or a parasite. Thus, fungal endo                α-1,4-D-polygalacturonases facilitate fungal                colonization and plant nutrient release by solubilizing                plant cell wall homo-α-1,4-D-galacturonate. See Lamb et                al., Bio/Technology 10: 1436 (1992). The cloning and                characterization of a gene that encodes a bean                endopolygalacturonase-inhibiting protein is described by                Toubart et al., Plant J. 2: 367 (1992).            -   xviii. A developmental-arrestive protein produced in                nature by a plant. For example, Logemann et al.,                Bio/Technology 10: 305 (1992), have shown that                transgenic plants expressing the barley                ribosome-inactivating gene have an increased resistance                to fungal disease.        -   b. Genes that Confer Resistance to a Herbicide, for Example:            -   i. A herbicide that inhibits the growing point or                meristem, such as an imidazalinone or a sulfonylurea.                Exemplary genes in this category code for mutant ALS and                AHAS enzyme as described, for example, by Lee et al.,                EMBO J. 7: 1241 (1988), and Miki et al., Theor. Appl.                Genet. 80: 449 (1990), respectively.            -   ii. Glyphosate (resistance imparted by mutant                5-enolpyruvl-3-phosphikimate synthase (EPSP) and aroA                genes, respectively) and other phosphono compounds such                as glufosinate (phosphinothricin acetyl transferase                (PAT) and Streptomyces hygroscopicus phosphinothricin                acetyl transferase (bar) genes), and pyridinoxy or                phenoxy proprionic acids and cyclohexones (ACCase                inhibitor-encoding genes). See, for example, U.S. Pat.                No. 4,940,835 to Shah et al., which discloses the                nucleotide sequence of a form of EPSP that can confer                glyphosate resistance. A DNA molecule encoding a mutant                aroA gene can be obtained under ATCC accession No.                39256, and the nucleotide sequence of the mutant gene is                disclosed in U.S. Pat. No. 4,769,061 to Comai. European                patent application No. 0 333 033 to Kumada et al. and                U.S. Pat. No. 4,975,374 to Goodman et al. disclose                nucleotide sequences of glutamine synthetase genes that                confer resistance to herbicides such as                L-phosphinothricin. The nucleotide sequence of a                phosphinothricin-acetyl-transferase gene is provided in                European application No. 0 242 246 to Leemans et al.                Furthermore, De Greef et al., Bio/Technology 7: 61                (1989), describe the production of transgenic plants                that express chimeric bar genes coding for                phospinothricin acetyl transferase activity. Exemplary                of genes conferring resistance to phenoxy proprionic                acids and cyclohexones, such as sethoxydim and                haloxyfop, are the Acc1-S1, Acc1-S2 and Acc1-S3 genes                described by Marshall et al., Theor. Appl. Genet. 83:                435 (1992).            -   iii. A herbicide that inhibits photosynthesis, such as a                triazine (psbA and gs+ genes) and a benzonitrile                (nitrilase gene). Przibilla et al., Plant Cell 3: 169                (1991), describe the transformation of Chlamydomonas                with plasmids encoding mutant psbA genes. Nucleotide                sequences for nitrilase genes are disclosed in U.S. Pat.                No. 4,810,648 to Stalker, and DNA molecules containing                these genes are available under ATCC Accession Nos.                53435, 67441 and 67442. Cloning and expression of DNA                coding for a glutathione S-transferase is described by                Hayes et al., Biochem. J. 285: 173 (1992).        -   c. Genes that Confer or Contribute to a Value-added Trait,            such as:            -   i. Modified fatty acid metabolism, for example, by                transforming a plant with an antisense gene of                stearoyl-ACP desaturase to increase stearic acid content                of the plant. See Knultzon et al., Proc. Natl. Acad.                Sci. USA 89: 2624 (1992).            -   ii. Decreased phytate content:                -   (i) Introduction of a phytase-encoding gene would                    enhance breakdown of phytate, adding more free                    phosphate to the transformed plant. For example, see                    Van Hartingsveldt et al., Gene 127: 87 (1993), for a                    disclosure of the nucleotide sequence of an                    Aspergillus niger phytase gene.                -   (ii) A gene could be introduced that reduces phytate                    content. In cotton, this, for example, could be                    accomplished, by cloning and then reintroducing DNA                    associated with the single allele that is                    responsible for cotton mutants characterized by low                    levels of phytic acid. See Raboy et al., Maydica 35:                    383 (1990).                -   (iii) Modified carbohydrate composition effected,                    for example, by transforming plants with a gene                    coding for an enzyme that alters the branching                    pattern of starch. See Shiroza et al., J. Bacteriol.                    170: 810 (1988) (nucleotide sequence of                    Streptococcus mutans fructosyltransferase gene),                    Steinmetz et al., Mol. Gen. Genet. 200: 220 (1985)                    (nucleotide sequence of Bacillus subtillus                    levansucrase gene), Pen et al., Bio/Technology 10:                    292 (1992) (production of transgenic plants that                    express Bacillus licheniformis α-amylase), Elliot et                    al., Plant Molec. Biol. 21: 515 (1993) (nucleotide                    sequences of tomato invertase genes), Sogaard et                    al., J. Biol. Chem. 268: 22480 (1993) (site-directed                    mutagenesis of barley α-amylase gene), and Fisher et                    al., Plant Physiol. 102: 1045 (1993) (corn endosperm                    starch branching enzyme II).

    -   E. Methods for Cotton Transformation

Numerous methods for plant transformation have been developed, includingbiological and physical, plant transformation protocols. See, forexample, Miki et al., “Procedures for Introducing Foreign DNA intoPlants” in Methods in Plant Molecular Biology and Biotechnology, Glick,B. R. and Thompson, J. E. Eds. (CRC Press, Inc., Boca Raton, 1993) pages67-88. In addition, expression vectors and in vitro culture methods forplant cell or tissue transformation and regeneration of plants areavailable. See, for example, Gruber et al., “Vectors for PlantTransformation” in Methods in Plant Molecular Biology and Biotechnology,Glick, B. R. and Thompson, J. E. Eds. (CRC Press, inc., Boca Raton,1993) pages 89-119.

-   -   -   a. Agrobacterium-Mediated Transformation

One method for introducing an expression vector into plants is based onthe natural transformation system of Agrobacterium. See, for example,Horsch et al., Science 227: 1229 (1985). A. tumefaciens and A.rhizogenes are plant pathogenic soil bacteria that genetically transformplant cells. The Ti and Ri plasmids of A. tumefaciens and A. rhizogenes,respectively, carry genes responsible for genetic transformation of theplant. See, for example, Kado, C. I., Crit. Rev. Plant. Sci.10: 1(1991). Descriptions of Agrobacterium vector systems and methods forAgrobacterium-mediated gene transfer are provided by Gruber et al.,supra, Mild et al., supra, and Moloney et al., Plant Cell Reports 8: 238(1989). See also, U.S. Pat. No. 5,591,616, issued Jan. 7, 1997.

-   -   -   2. Direct Gene Transfer

Several methods of plant transformation, collectively referred to asdirect gene transfer, have been developed as an alternative toAgrobacterium-mediated transformation.

A generally applicable method of plant transformation ismicroprojectile-mediated transformation wherein DNA is carried on thesurface of microprojectiles measuring 1 to 4 pm (See e.g., U.S. Pat. No.5,550,318; U.S. Pat. No. 5,736,369; U.S. Pat. No. 5,538,880; and PCTPublication WO 95/06128). The expression vector is introduced into planttissues with a biolistic device that accelerates the microprojectiles tospeeds of 300 to 600 m/s, which is sufficient to penetrate plant cellwalls and membranes. Sanford et al, Part. Sci. Technol. 5: 27 (1987),Sanford, J. C., Trends Biotech. 6: 299 (1988), Klein et al.,Bio/Technology 6: 559-563 (1988), Sanford, J. C., Physiol Plant 79: 206(1990), Klein et al., Biotechnology 10: 268 (1992).

Another method for physical delivery of DNA to plants is sonication oftarget cells. Zhang et al., Bio/Technology 9: 996 (1991). Alternatively,liposome or spheroplast fusion have been used to introduce expressionvectors into plants. Deshayes et al., EMBO J., 4: 2731 (1985), Christouet al., Proc. Natl. Acad. Sci. U.S.A. 84: 3962 (1987). Direct uptake ofDNA into protoplasts using CaCl₂ precipitation, polyvinyl alcohol orpoly-L-ornithine have also been reported. Hain et al., Mol. Gen. Genet.199: 161 (1985) and Draper et al., Plant Cell Physiol. 23: 451 (1982).Electroporation of protoplasts and whole cells and tissues have alsobeen described. U.S. Pat. No. 5,384,253 and Donn et al., in Abstracts ofVIIth International Congress on Plant Cell and Tissue Culture IAPTC,A2-38, p 53 (1990); D'Halluin et al., Plant Cell 4: 1495-1505 (1992) andSpencer et al., Plant Mol. Biol. 24: 51-61 (1994).

Other methods that have been described for the genetic transformation ofcotton include electrotransformation (U.S. Pat. No. 5,371,003) andsilicon carbide fiber-mediated transformation (U.S. Pat. No. 5,302,532and U.S. Pat. No. 5,464,765).

Following transformation of cotton target tissues, expression of theabove-described selectable marker genes allows for preferentialselection of transformed cells, tissues and/or plants, usingregeneration and selection methods now well known in the art.

The foregoing methods for transformation are typically used forproducing transgenic cotton varieties. Transgenic cotton varieties canthen be crossed, with another (non-transformed or transformed) cottonvariety, to produce a transgenic hybrid cotton plant. Alternatively, agenetic trait that has been engineered into a particular cotton varietyusing the foregoing transformation techniques can be moved into anotherline using traditional backcrossing techniques that are well known inthe plant breeding arts. For example, a backcrossing approach can beused to move an engineered trait from a public, non-elite line into anelite line, or from a hybrid cotton plant containing a foreign gene inits genome into a line or lines that do not contain that gene.

IX. Genetic Complements

In addition to phenotypic observations, a plant can also be described byits genotype. The genotype of a plant can be described through a geneticmarker profile that can identify plants of the same variety, a relatedvariety or be used to determine or to validate a pedigree. Geneticmarker profiles can be obtained by techniques such as RestrictionFragment Length Polymorphisms (RFLPs), Randomly Amplified PolymorphicDNAs (RAPDs), Arbitrarily Primed Polymerase Chain Reaction (AP-PCR), DNAAmplification Fingerprinting (DAF), Sequence Characterized AmplifiedRegions (SCARs), Amplified Fragment Length Polymorphisms (AFLPs), SimpleSequence Repeats (SSRs), which are also referred to as Microsatellites,and Single Nucleotide Polymorphisms (SNPs), Isozyme Electrophoresis andIsoelectric Focusing.

Particular markers used for these purposes are not limited to the set ofmarkers disclosed herewithin, but are envisioned to include any type ofgenetically stable marker and marker profile that provides a means ofdistinguishing varieties. In addition to being used for identificationof cotton varieties, a hybrid produced through the use of P07X.8212.RF,and identification or verification of the pedigree of progeny plantsproduced through the use of P07X.8212.RF, the genetic marker profile isalso useful in breeding and developing backcross conversions.

Means of performing genetic marker profiles using SSR polymorphisms arewell known in the art. SSRs are genetic markers based on polymorphismsin repeated nucleotide sequences, such as microsatellites. The phrase“simple sequence repeats” or “SSR” refers to di-, tri- or tetra-nucleotide repeats within a genome. The repeat region may vary in lengthbetween genotypes while the DNA flanking the repeat is conserved suchthat the primers will work in a plurality of genotypes. A polymorphismbetween two genotypes represents repeats of different lengths betweenthe two flanking conserved DNA sequences. A marker system based on SSRscan be highly informative in linkage analysis relative to other markersystems in that multiple alleles may be present. Another advantage ofthis type of marker is that, through use of flanking primers, detectionof SSRs can be achieved, for example, by the polymerase chain reaction(PCR). The PCR detection is done by the use of two oligonucleotideprimers flanking the polymorphic segment of repetitive DNA followed byDNA amplification. This step involves repeated cycles of heatdenaturation of the DNA followed by annealing of the primers to theircomplementary sequences at low temperatures, and extension of theannealed primers with DNA polymerase. Size separation of DNA fragmentson agarose or polyacrylamide gels following amplification comprises themajor part of the methodology.

DNA isolation and amplification may be performed in certain embodimentsof the present disclosure as follows. DNA may be extracted from plantleaf tissue using DNeasy 96 Plant Kit from Qiagen, Inc. (Valencia,Calif., U.S.A.) following an optimized September 2002 manufacturer'sprotocol. PCR amplifications are performed using a Qiagen HotStar™ Taqmaster mix in an 8 μl reaction format as follows: 2 μl DNA (5 ng/μL+6 μLof master mix). The PCR conditions are as follows: 12 mins. at 95° C.,40 cycles of 5 seconds at 94° C., 15 seconds at 55° C., 30 seconds at72° C., 30 mins at 72° C., followed by cooling to 4° C. Followingisolation and amplification, markers can be scored by gelelectrophoresis of the amplification products. Scoring of markergenotype is based on the size of the amplified fragment as measured bymolecular weight (MW) rounded to the nearest integer. Multiple samples,comprised of fluorescently labeled DNA fragments may be processed in anABI 3700 capillary-based machine and precise allele sizing and locusgenotyping can be done by running GeneScan and Genotyper software (PEApplied Biosystems, Foster City, Calif.). When comparing varieties, itis preferable that all SSR profiles be performed in the same lab. An SSRservice is available to the public on a contractual basis by Paragen,Research Triangle Park, N.C. (formerly Celera AgGen of Davis, Calif.).

All publications, patents and patent applications mentioned in thespecification are indicative of the level of those skilled in the art towhich this disclosure pertains. All such publications, patents andpatent applications are incorporated by reference herein to the sameextent as if each was specifically and individually indicated to beincorporated by reference herein.

The foregoing invention has been described in some detail by way ofillustration and example for purposes of clarity and understanding.However, it should be appreciated by those having ordinary skill in theart that certain changes and modifications such as single genemodifications and mutations, somoclonal variants, variant individualsselected from large populations of the plants of the instant variety andthe like may be practiced within the scope of the embodiments of theinvention, as limited only by the scope of the appended claims, withoutdeparting from the true concept, spirit, and scope of the invention.

1. A seed of cotton variety designated P07X.8212.RF, or a part thereof,representative seed of the variety having been deposited under ATCCAccession No. PTA-11344.
 2. The seed part of claim 1 selected from thegroup consisting of hull (seedcoat), germ and endosperm.
 3. The seed ofclaim 1, further comprising a coating.
 4. A substantially homogenouscomposition of the cotton seed of claim
 1. 5. A method for producing aseed of a cotton plant, comprising: (a) planting seed of claim 1 inproximity to itself or to different seed from a same variety; (b)growing plants from the seed under pollinating conditions; and, (c)harvesting resultant seed.
 6. A cotton seed produced by the method ofclaim
 5. 7. The method of claim 5, further comprising pre-treating theseed of claim 1 before performing step (a).
 8. The method of claim 5,further comprising treating the growing plants or soil surrounding thegrowing plants with an agricultural chemical.
 9. A cotton plant producedby growing the seed of claim
 1. 10. A part of the cotton plant of claim9, selected from the group consisting of an intact plant cell, a plantprotoplast, embryos, pollen, flowers, seeds, linters, fibers, pods,gossypol glands, leaves, bolls, stems, roots, root tips, and anthers.11. Fibers of the plant of claim
 9. 12. Staples of the plant of claim 9.13. A cotton plant, or a part thereof, having all the physiological andmorphological characteristics of the cotton plant of claim
 9. 14. Asubstantially homogenous population of cotton plants of claim
 9. 15. Thesubstantially homogenous population of cotton plants of claim 14,wherein the population is present in a field and the field furthercomprises other, different cotton plants.
 16. A method for producing acotton plant, comprising: (a) crossing cotton variety plantP07X.8212.RF, representative seed of the cultivar having been depositedunder ATCC Accession No. PTA-11344, with another different cotton plantto yield progeny cotton seed.
 17. The method of claim 16, wherein theother, different cotton plant is a cotton variety.
 18. The method ofclaim 16, further comprising: (b) growing the progeny cotton seed fromstep (a) under self-pollinating or sib-pollinating conditions for about5 to about 7 generations; and (c) harvesting resultant seed.
 19. Themethod of claim 16, further comprising selecting plants obtained fromgrowing at least one generation of the progeny cotton seed for adesirable trait.
 20. A method of introducing a desired trait into cottonvariety P07X.8212.RF, representative seed of the variety having beendeposited under ATCC Accession No. PTA-11344, comprising: (a) crossingP07X.8212.RF plants with plants of another cotton variety that comprisea desired trait to produce F₁ progeny plants; (b) selecting F₁ progenyplants that have the desired trait; (c) crossing selected progeny plantswith P07X.8212.RF plants to produce backcross progeny plants; (d)selecting for backcross progeny plants that comprise the desired traitand physiological and morphological characteristics of cotton varietyP07X.8212.RF; and (e) performing steps (c) and (d) one or more times insuccession to produce the selected or higher backcross progeny plantsthat comprise the desired trait and all of the physiological andmorphological characteristics of cotton variety P07X.8212.RF listed inTable 1 as determined at the 5% significance level when grown in thesame environmental conditions.